Tetrahydropyranyl cyclopentyl tetrahydropyridopyridine modulators of chemokine receptor activity

ABSTRACT

The present invention is directed to compounds of the formula I: 
                         
(wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , X, n and the dashed line are defined herein) which are useful as modulators of chemokine receptor activity. In particular, these compounds are useful as modulators of the chemokine receptor CCR-2. The present invention is also directed to intermediates useful in the preparation of formula I compounds.

This application claims benefit as a CIP of PCT/US03/13042, filed Apr.25, 2003, which claims benefit of U.S. Provisional Application60/376,291, filed Apr. 29, 2002.

BACKGROUND OF THE INVENTION

The chemokines are a family of small (70-120amino acids),proinflammatory cytokines, with potent chemotactic activities.Chemokines are chemotactic cytokines that are released by a wide varietyof cells to attract various cells, such as monocytes, macrophages, Tcells, eosinophils, basophils and neutrophils to sites of inflammation(reviewed in Schall, Cytokine, 3, 165-183(1991) and Murphy, Rev. Immun.,12, 593-633(1994)). These molecules were originally defined by fourconserved cysteines and divided into two subfamilies based on thearrangement of the first cysteine pair. In the CXC-chemokine family,which includes IL-8, GROα, NAP-2 and IP-10, these two cysteines areseparated by a single amino acid, while in the CC-chemokine family,which includes RANTES, MCP-1, MCP-2, MCP-3, MIP-1α, MIP-1β and eotaxin,these two residues are adjacent.

The α-chemokines, such as interleukin-8 (IL-8), neutrophil-activatingprotein-2 (NAP-2) and melanoma growth stimulatory activity protein(MGSA) are chemotactic primarily for neutrophils, whereas β-chemokines,such as RANTES, MIP-1α, MIP-1β, monocyte chemotactic protein-1 (MCP-1),MCP-2, MCP-3 and eotaxin are chemotactic for macrophages, monocytes,T-cells, eosinophils and basophils (Deng, et al., Nature, 381, 661-666(1996)).

The chemokines are secreted by a wide variety of cell types and bind tospecific G-protein coupled receptors (GPCRs) (reviewed in Horuk, TrendsPharm. Sci., 15, 159-165 (1994)) present on leukocytes and other cells.These chemokine receptors form a sub-family of GPCRs, which, at present,consists of fifteen characterized members and a number of orphans.Unlike receptors for promiscuous chemoattractants such as C5a, fMLP,PAF, and LTB4, chemokine receptors are more selectively expressed onsubsets of leukocytes. Thus, generation of specific chemokines providesa mechanism for recruitment of particular leukocyte subsets.

On binding their cognate ligands, chemokine receptors transduce anintracellular signal though the associated trimeric G protein, resultingin a rapid increase in intracellular calcium concentration. There are atleast seven human chemokine receptors that bind or respond toα-chemokines with the following characteristic pattern: CCR-1 (or“CKR-1” or “CC-CKR-1”) [MIP-1α, MIP-1β, MCP-3, RANTES] (Ben-Barruch, etal., J. Biol. Chem., 270, 22123-22128 (1995); Beote, et al, Cell, 72,415-425 (1993)); CCR-2A and CCR-2B (or “CKR-2A”/“CKR-2A” or“CC-CKR-2A”/“CC-CKR-2A”) [MCP-1, MCP-2, MCP-3, MCP-4]; CCR-3(or “CKR-3”or “CC-CKR-3”) [Eotaxin, Eotaxin 2, RANTES, MCP-2, MCP-3] (Rollins, etal., Blood, 90, 908-928(1997)); CCR-4 (or “CKR-4” or “CC-CKR-4”)[MIP-1α,RANTES, MCP-1] (Rollins, et al., Blood, 90, 908-928 (1997)); CCR-5 (or“CKR-5” or “CC-CKR-5”) [MIP-1α, RANTES, MIP-1β] (Sanson, et al.,Biochemistry, 35, 3362-3367 (1996)); and the Duffy blood-group antigen[RANTES, MCP-1] (Chaudhun, et al., J. Biol. Chem., 269, 7835-7838(1994)). The β-chemokines include eotaxin, MIP (“macrophage inflammatoryprotein”), MCP (“monocyte chemoattractant protein”) and RANTES(“regulation-upon-activation, normal T expressed and secreted”) amongother chemokines.

Chemokine receptors, such as CCR-1, CCR-2, CCR-2A, CCR-2B, CCR-3, CCR-4,CCR-5, CXCR-3, CXCR-4, have been implicated as being important mediatorsof inflammatory and immunoregulatory disorders and diseases, includingasthma, rhinitis and allergic diseases, as well as autoimmunepathologies such as rheumatoid arthritis and atherosclerosis. Humans whoare homozygous for the 32-basepair deletion in the CCR-5 gene appear tohave less susceptibility to rheumatoid arthritis (Gomez, et al.,Arthritis & Rheumatism, 42, 989-992 (1999)). A review of the role ofeosinophils in allergic inflammation is provided by Kita, H., et al., J.Exp. Med. 183, 2421-2426 (1996). A general review of the role ofchemokines in allergic inflammation is provided by Lustger, A. D., NewEngland J. Med., 338(7), 426-445 (1998).

A subset of chemokines are potent chemoattractants for monocytes andmacrophages. The best characterized of these is MCP-1 (monocytechemoattractant protein-1), whose primary receptor is CCR2. MCP-1 isproduced in a variety of cell types in response to inflammatory stimuliin various species, including rodents and humans, and stimulateschemotaxis in monocytes and a subset of lymphocytes. In particular,MCP-1 production correlates with monocyte and macrophage infiltration atinflammatory sites. Deletion of either MCP-1 or CCR2 by homologousrecombination in mice results in marked attenuation of monocyterecruitment in response to thioglycollate injection and Listeriamonocytogenes infection (Lu et al., J. Exp. Med., 187, 601-608 (1998);Kurihara et al. J. Exp. Med., 186, 1757-1762 (1997); Boring et al. J.Clin. Invest., 100, 2552-2561 (1997); Kuziel et al. Proc. Natl. Acad.Sci., 94, 12053-12058 (1997)). Furthermore, these animals show reducedmonocyte infiltration into granulomatous lesions induced by theinjection of schistosomal or mycobacterial antigens (Boring et al. J.Clin. Invest., 100, 2552-2561 (1997); Warmington et al. Am J. Path.,154, 1407-1416 (1999)). These data suggest that MCP-1-induced CCR2activation plays a major role in monocyte recruitment to inflammatorysites, and that antagonism of this activity will produce a sufficientsuppression of the immune response to produce therapeutic benefits inimmunoinflammatory and autoimmune diseases.

Accordingly, agents which modulate chemokine receptors such as the CCR-2receptor would be useful in such disorders and diseases.

In addition, the recruitment of monocytes to inflammatory lesions in thevascular wall is a major component of the pathogenesis of atherogenicplaque formation. MCP-1 is produced and secreted by endothelial cellsand intimal smooth muscle cells after injury to the vascular wall inhypercholesterolemic conditions. Monocytes recruited to the site ofinjury infiltrate the vascular wall and differentiate to foam cells inresponse to the released MCP-1. Several groups have now demonstratedthat aortic lesion size, macrophage content and necrosis are attenuatedin MCP-1 −/− or CCR2 −/− mice backcrossed to APO-E −/−, LDL-R −/− or ApoB transgenic mice maintained on high fat diets (Boring et al. Nature,394, 894-897 (1998); Gosling et al. J. Clin. Invest., 103, 773-778(1999)). Thus, CCR2 antagonists may inhibit atherosclerotic lesionformation and pathological progression by impairing monocyte recruitmentand differentiation in the arterial wall.

SUMMARY OF THE INVENTION

The present invention is directed to compounds which are modulators ofchemokine receptor activity and are useful in the prevention ortreatment of certain inflammatory and immunoregulatory disorders anddiseases, allergic diseases, atopic conditions including allergicrhinitis, dermatitis, conjunctivitis, and asthma, as well as autoimmunepathologies such as rheumatoid arthritis and atherosclerosis. Theinvention is also directed to pharmaceutical compositions comprisingthese compounds and the use of these compounds and compositions in theprevention or treatment of such diseases in which chemokine receptorsare involved. Further, the present invention is directed to theintermediate compounds useful in the preparation of these chemokinereceptor modulators.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to compounds of the formula I:

Iwherein:

-   X is selected from the group consisting of:    -   —O—, —NR²⁰ —, —S—, —SO—, —SO₂ —, and —CR²¹R²² —, —NSO₂ R²⁰ —,        —NCOR²⁰ —, —NCO₂ R²⁰ —, —CR²¹CO₂ R²⁰ —, —CR²¹OCOR²⁰ —, —CO—,        where R²⁰ is selected from: hydrogen, C₁₋₆ alkyl, benzyl,        phenyl, C₃₋₆ cycloalkyl where the alkyl, phenyl, benzyl, and        cycloalkyl groups can be unsubstituted or substituted with 1-3        substituents where the substituents are independently selected        from: halo, hydroxy, C₁₋₃alkyl, C₁₋₃alkoxy, —CO₂H,        —CO₂—C₁₋₆alkyl, and trifluoromethyl,    -   where R²¹ and R²² are independently selected from: hydrogen,        hydroxy, C₁₋₆ alkyl, —O—C₁₋₆ alkyl, benzyl, phenyl, C₃₋₆        cycloalkyl where the alkyl, phenyl, benzyl, and cycloalkyl        groups can be unsubstituted or substituted with 1-3 substituents        where the substituents are independently selected from: halo,        hydroxy, C₁₋₃ alkyl, C₁₋₃ alkoxy, —CO₂ H, —CO₂ —C₁₋₆ alkyl, and        trifluoromethyl;-   R¹ is selected from:    -   —C₁₋₆ alkyl, —C₀₋₆ alkyl-O—C₁₋₆ alkyl-, —C₀₋₆ alkyl-S—C₁₋₆        alkyl-, —(C₀₋₆alkyl)-(C₃₋₇cycloalkyl)-(C₀₋₆alkyl), hydroxy, —CO₂        R²⁰, heterocycle, —CN, —NR²⁰ R²⁶ —, —NSO₂ R²⁰ —, —NCOR²⁰ —,        —NCO₂ R²⁰ —, —NCOR²⁰ —, —CR²¹CO₂R²⁰—, —CR²¹OCOR²⁰—, phenyl and        pyridyl,    -   where R²⁶ is selected from: hydrogen, C₁₋₆ alkyl, benzyl,        phenyl, C₃₋₆ cycloalkyl where the alkyl, phenyl, benzyl, and        cycloalkyl groups can be unsubstituted or substituted with 1-3        substituents where the substituents are independently selected        from: halo, hydroxy, C₁₋₃alkyl, C₁₋₃alkoxy, —CO₂H, —CO₂—C₁₋₆        alkyl, and trifluoromethyl    -   where the alkyl and the cycloalkyl are unsubstituted or        substituted with 1-7substituents where the substituents are        independently selected from:        -   (a) halo,        -   (b) hydroxy,        -   (c) —O—C₁₋₃alkyl,        -   (d) trifluoromethyl,        -   (f) C₁₋₃alkyl,        -   (g) —O—C₁₋₃alkyl,        -   (h) —CO₂R²⁰,        -   (i) —SO₂R²⁰,        -   (j) —NHCOCH₃,        -   (k) —NHSO₂CH₃,        -   (l) -heterocycle,        -   (m) ═O,        -   (n) —CN,            and where the phenyl and pyridyl are unsubstituted or            substituted with 1-3 substituents where the substituents are            independently selected from: halo, hydroxy, C₁₋₃ alkyl, C₁₋₃            alkoxy and trifluoromethyl;-   R² is selected from:    -    (a) hydrogen,    -    (b) hydroxy,    -    (c) halo,    -    (d) C₁₋₃ alkyl, where the alkyl is unsubstituted or substituted        with 1-6 substituents independently selected from: fluoro, and        hydroxy,    -    (e) —NR²⁰R²⁶,    -    (f) —CO₂R²⁰,    -    (g) —CONR²⁰R²⁶,    -    (h) —NR²⁰COR²¹,    -    (i) —OCONR²⁰R²⁶,    -    (j) —NR²⁰CONR²⁰R²⁶,    -    (k) -heterocycle,    -    (l) —CN,    -    (m) —NR²⁰—SO₂—NR²⁰R²⁶,    -    (n) —NR²⁰—SO₂—R²⁶,    -    (O) —SO₂—NR²⁰R²⁶, and    -    (p) ═O, where R² is connected to the ring via a double bond;-   R³ is oxygen or is absent;-   R⁴ is selected from:    -    (a) hydrogen,    -    (b) C₁₋₆ alkyl,    -    (c) trifluoromethyl,    -    (d) trifluoromethoxy,    -    (e) chloro,    -    (f) fluoro,    -    (g) bromo, and    -    (h) phenyl;-   R⁵ is selected from:    -    (a) C₁₋₆alkyl, where alkyl may be unsubstituted or substituted        with 1-6 fluoro and optionally substituted with hydroxyl,    -    (b) —O—C₁₋₆alkyl, where alkyl may be unsubstituted or        substituted with 1-6 fluoro,    -    (c) —CO—C₁₋₆alkyl, where alkyl may be unsubstituted or        substituted with 1-6 fluoro,    -    (d) —S—C₁₋₆alkyl, where alkyl may be unsubstituted or        substituted with 1-6 fluoro,    -    (e)-pyridyl, which may be unsubstituted or substituted with one        or more substituents selected from the group consisting of:        halo, trifluoromethyl, C₁₋₄alkyl, and CO₂R²⁰,    -    (f) fluoro,    -    (g) chloro,    -    (h) bromo,    -    (i) —C₄₋₆cycloalkyl,    -    (j) —O—C₄₋₆cycloalkyl,    -    (k) phenyl, which may be unsubstituted or substituted with one        or more substituents selected from the group consisting of:        halo, trifluoromethyl, C₁₋₄alkyl, and CO₂R²⁰,    -    (l) —O-phenyl, which may be unsubstituted or substituted with        one or more substituents selected from the group consisting of:        halo, trifluoromethyl, C₁₋₄alkyl, and CO₂R²⁰,    -    (m) —C₃₋₆cycloalkyl, where alkyl may be unsubstituted or        substituted with 1-6 fluoro,    -    (n) —O—C₃₋₆cycloalkyl, where alkyl may be unsubstituted or        substituted with 1-6 fluoro,    -    (O)-heterocycle,    -    (p) —CN, and    -    (q) —CO₂R²⁰;-   R⁶ is selected from:    -    (a) hydrogen,    -    (b) C₁₋₆alkyl, and    -    (c) trifluoromethyl    -    (d) fluoro    -    (e) chloro, and    -    (f) bromo;-   R⁷ is selected from:    -    (a) hydrogen, and    -    (b) C₁₋₆alkyl, which is unsubstituted or substituted with 1-3        substituents where the substituents are independently selected        from: halo, hydroxy, —CO₂H, —CO₂C₁₋₆alkyl, and —O—C₁₋₃alkyl;-   R⁸ is selected from:    -    (a) hydrogen,    -    (b) C₁₋₆alkyl, where alkyl may be unsubstituted or substituted        with 1-6 substituents where the substituents are chosen from the        group: fluoro, C₁₋₃alkoxy, hydroxy, —CO₂R²⁰,    -    (c) fluoro,    -    (d) —O—C₁₋₃alkyl, where alkyl may be unsubstituted or        substituted with 1-3 fluoro, and    -    (e) C₃₋₆ cycloalkyl,    -    (f) —O—C₃₋₆cycloalkyl,    -    (g) hydroxy,    -    (h) —CO₂R²⁰,    -    (i) —OCOR²⁰,    -    or R⁷ and R⁸ may be joined together via a C₂₋₄alkyl or a        C₀₋₂alkyl-O—C₁₋₃alkyl chain to form a 5-7 membered ring;-   R⁹ is selected from:    -    (a) hydrogen,    -    (b) C₁₋₆alkyl, where alkyl may be unsubstituted or substituted        with 1-6 substituents where the substituents are chosen from the        group: fluoro, C₁₋₃alkoxy, hydroxy, —CO₂R²⁰,    -    (c) CO₂R²⁰,    -    (d) hydroxy, and    -    (e) —O—C₁₋₆alkyl, where alkyl may be unsubstituted or        substituted with 1-6 substituents where the substituents are        chosen from the group: fluoro, C₁₋₃alkoxy, hydroxy, —CO₂R²⁰,    -    or R⁸ and R⁹ may be joined together by a C₁₋₄alkyl chain or a    -    C₀₋₃alkyl-O—C₀₋₃alkyl chain to form a 3-6 membered ring;-   R¹⁰ is selected from:    -    (a) hydrogen, and    -    (b) C₁₋₆alkyl, where alkyl may be unsubstituted or substituted        with 1-6 fluoro,    -    (c) fluoro,    -    (d) —O—C₃₋₆cycloalkyl, and    -    (e) —O—C₁₋₃alkyl, where alkyl may be unsubstituted or        substituted with 1-6 fluoro, or R⁸ and R¹⁰ may be joined        together by a C₂₋₃alkyl chain to form a 5-6 membered ring, where        the alkyl are unsubstituted or substituted with 1-3 substituents        where the substiuents are independently selected from: halo,        hydroxy, —CO₂R²⁰, C₁₋₃alkyl, and C₁₋₃alkoxy,    -    or R⁸ and R¹⁰ may be joined together by a C₁₋₂alkyl-O—C₁₋₂alkyl        chain to form a 6-8 membered ring, where the alkyl are        unsubstituted or substituted with 1-3 substituents where the        substiuents are independently selected from: halo, hydroxy,        —CO₂R²⁰, C₁₋₃alkyl, and    -    C₁₋₃ alkoxy,    -    or R⁸ and R¹⁰ may be joined together by a —O—C₁₋₂alkyl-O-chain        to form a 6-7 membered ring, where the alkyl are unsubstituted        or substituted with 1-3 substituents where the substiuents are        independently selected from: halo, hydroxy, —CO₂R²⁰, C₁₋₃alkyl,        and    -    C₁₋₃alkoxy;-   n is selected from 0, 1 and 2;-   the dashed line represents a single or a double bond;-   and pharmaceutically acceptable salts thereof and individual    diastereomers thereof.

Preferred compounds of the present invention include those of formulaIa:

wherein R¹, R², R³, R⁵, R⁷, R⁸, R¹⁰ and X are defined herein,

-   and pharmaceutically acceptable salts and individual diastereomers    thereof.

More preferred compounds of the present invention also include those offormula Ib:

wherein R¹, R², R³, R⁵, R⁸ and R¹⁰ are defined herein, andpharmaceutically acceptable salts and individual diastereomers thereof.

Even more preferred compounds of the present invention also includethose of formula Ic:

wherein R¹, R³, R⁵ and R⁸ are defined herein.

Still more preferred compounds of the present invention also includethose of formula Id:

wherein R³ and R⁸ are defined herein and pharmaceutically acceptablesalts and individual diastereomers thereof.

In the present invention it is preferred that X is selected from thegroup consisting of:

-   -   —O—, —CH₂—, —S—, —SO—, and —SO₂—.

In the present invention it is more preferred that X is selected fromthe group consisting of: —O—, and —CH₂—.

In the present invention it is even more preferred that X is —O—.

In the present invention it is preferred that R¹ is selected from:

-   -   (1) —C₁₋₆alkyl, which is unsubstituted or substituted with 1-6        substituents where the substituents are independently selected        from:        -   (a) halo,        -   (b) hydroxy,        -   (c) —O—C₁₋₃alkyl, and        -   (d) trifluoromethyl,    -   (2) —C₀₋₆alkyl-O—C₁₋₆alkyl-, which is unsubstituted or        substituted with 1-6 substituents where the substituents are        independently selected from:        -   (a) halo, and        -   (b) trifluoromethyl,    -   (3) —C₀₋₆alkyl-S—C₁₋₆alkyl-, which is unsubstituted or        substituted with 1-6 substituents where the substituents are        independently selected from:        -   (a) halo, and        -   (b) trifluoromethyl,    -   (4) —(C₃₋₅cycloalkyl)-(C₀₋₆alkyl), which is unsubstituted or        substituted with 1-7 substituents where the substituents are        independently selected from:        -   (a) halo,        -   (b) hydroxy,        -   (c) —O—C₁₋₃alkyl, and        -   (d) trifluoromethyl.

In the present invention it is more preferred that R¹ is C₁₋₆alkyl whichis unsubstituted or substituted with 1-5substituents where thesubstituents are independently selected from:

-   -   -   (a) hydroxy, and        -   (b) fluoro.

In the present invention it is even more preferred that R¹ is selectedfrom:

-   -   -   (a) isopropyl,        -   (b) —CH(OH)CH₃, and        -   (c) —CH₂CF₃.

In the present invention it is still more preferred that R¹ isisopropyl.

In the present invention it is preferred that R² is selected from:

-   -   -   (a) hydrogen,        -   (b) hydroxy,        -   (c) —NH₂,        -   (d) —CO₂H,        -   (e) -triazolyl,        -   (f) -tetrazolyl,        -   (g) —CO₂—C₁₋₆alkyl,        -   (h) —CONH₂,        -   (i) —CONH—C₁₋₆alkyl,        -   (j) —NHCO—C₁₋₆alkyl,        -   (k) —NHCONH₂,        -   (l) —NHCONH—C₁₋₆alkyl        -   (m) —OCONH—C₁₋₆alkyl,        -   (n) —NH—SO₂—C₁₋₁₆alkyl, and        -   (O) —SO₂—NH—C₁₋₆alkyl.

In the present invention it is more preferred that R² is selected from:

-   -   -   (a) hydrogen,        -   (b) hydroxy,        -   (c) —NH₂,        -   (d) —CO₂H,        -   (e) -triazolyl,        -   (f) -tetrazolyl,        -   (g) —NHCOCH₃,        -   (h) —NHCONH₂,        -   (i) —CONH₂,        -   (j) —NH—SO₂—CH₃, and        -   (k) —SO₂—NH—CH₃.

In the present invention it is even more preferred that R² is hydrogen.

In the present invention it is preferred that R⁴ is selected from:

-   -   -   (a) hydrogen, and        -   (b) trifluoromethyl.

In the present invention it is more preferred that R⁴ is hydrogen.

In the present invention it is preferred that R⁵ is selected from:

-   -   -   (a) C₁₋₃alkyl substituted with 1-6 fluoro,        -   (b) chloro,        -   (c) bromo,        -   (d) —O-phenyl, which may be unsubstituted or substituted            with one or more substituents selected from the group            consisting of: halo and trifluoromethyl,        -   (e) phenyl, which may be unsubstituted or substituted with            one or more substituents selected from the group consisting            of: halo and trifluoromethyl, and        -   (f) —O—C₁₋₃alkyl substituted with 1-6 fluoro.

In the present invention it is more preferred that R⁵ is selected from:

-   -   -   (a) trifluoromethyl,        -   (b) trifluoromethoxy,        -   (c) bromo, and        -   (d) chloro.

In the present invention it is most preferred that R⁵ istrifluoromethyl.

In the present invention it is preferred that R⁶ is hydrogen.

In the present invention it is preferred that R⁷ is hydrogen or methyl.

In the present invention it is preferred that R⁸ is selected from:

-   -   -   (a) hydrogen,        -   (b) C₁₋₃alkyl, which is unsubstituted or substituted with            1-6 fluoro,        -   (c) —O—C₁₋₃alkyl, and        -   (d) fluoro, and        -   (e) hydroxy.

In the present invention it is more preferred that R⁸ is selected from:

-   -   -   (a) hydrogen,        -   (d) trifluoromethyl,        -   (c) methyl,        -   (d) methoxy,        -   (e) ethoxy,        -   (f) ethyl,        -   (g) fluoro, and        -   (h) hydroxy.

In the present invention it is preferred that R⁹ is hydrogen.

In the present invention it is preferred that R¹⁰ is selected from:

-   -   -   (a) hydrogen,        -   (b) methyl, and        -   (c) methoxy.

In the present invention it is preferred that R¹⁰ is hydrogen.

In the present invention it is also preferred that R⁸ and R¹⁰ are joinedtogether by a —CH₂CH₂— chain or a —CH₂CH₂CH₂— chain to form acyclopentyl ring or a cyclohexyl ring.

In the present invention it is preferred that n is 1.

Representative compounds of the present invention include thosepresented in the Examples and pharmaceutically acceptable salts andindividual diastereomers thereof.

The compounds of the instant invention have at least two asymmetriccenters at the 1- and 3-positions of the cyclopentyl ring and oneasymmetric center at the 4-position of the ring bearing X. Additionalasymmetric centers may be present depending upon the nature of thevarious substituents on the molecule. Each such asymmetric center willindependently produce two optical isomers and it is intended that all ofthe possible optical isomers and diastereomers in mixtures and as pureor partially purified compounds are included within the ambit of thisinvention. The absolute configurations of the more preferred compoundsof this orientation, where the substituents on the cyclopentyl ring(amide and amine units) are cis, as depicted:

The absolute configurations of the most preferred compounds of thisinvention are those of the orientation as depicted:

wherein the carbon bearing the amine substituent is designated as beingof the (R) absolute configuration and the carbon bearing the amidesubunit can be designated as being of either the (S) or (R) absoluteconfiguration depending on the priority for R¹. For example if R isisopropyl then the absolute stereochemistry at the carbon bearing theamide subunit would be (S) since the amide and amine units are preferredto have the cis arrangement on the cyclopentyl ring.

The independent syntheses of diastereomers and enantiomers or theirchromatographic separations may be achieved as known in the art byappropriate modification of the methodology disclosed herein. Theirabsolute stereochemistry may be determined by the x-ray crystallographyof crystalline products or crystalline intermediates which arederivatized, if necessary, with a reagent containing an asymmetriccenter of known absolute configuration.

As appreciated by those of skill in the art, halo or halogen as usedherein are intended to include chloro, fluoro, bromo and iodo.Similarly, C₁₋₈, as in C₁₋₈alkyl is defined to identify the group ashaving 1, 2, 3, 4, 5, 6, 7 or 8 carbons in a linear or branchedarrangement, such that C₁₋₈alkyl specifically includes methyl, ethyl,n-propyl, iso-propyl, n-butyl, iso-butyl, tert-butyl, pentyl, hexyl,heptyl and octyl. Likewise, C₀, as in C₀alkyl is defined to identify thepresence of a direct covalent bond. The term “heterocycle” as usedherein is intended to include the following groups: benzoimidazolyl,benzofuranyl, benzofurazanyl, benzopyrazolyl, benzotriazolyl,benzothiophenyl, benzoxazolyl, carbazolyl, carbolinyl, cinnolinyl,furanyl, imidazolyl, indolinyl, indolyl, indolazinyl, indazolyl,isobenzofuranyl, isoindolyl, isoquinolyl, isothiazolyl, isoxazolyl,naphthpyridinyl, oxadiazolyl, oxazolyl, oxetanyl, pyranyl, pyrazinyl,pyrazolyl, pyridazinyl, pyridopyridinyl, pyridazinyl, pyridyl,pyrimidyl, pyrrolyl, quinazolinyl, quinolyl, quinoxalinyl,tetrahydropyranyl, tetrazolyl, tetrazolopyridyl, thiadiazolyl,thiazolyl, thienyl, triazolyl, azetidinyl, 1,4-dioxanyl,hexahydroazepinyl, piperazinyl, piperidinyl, pyrrolidinyl, morpholinyl,thiomorpholinyl, dihydrobenzoimidazolyl, dihydrobenzofuranyl,dihydrobenzothiophenyl, dihydrobenzoxazolyl, dihydrofuranyl,dihydroimidazolyl, dihydroindolyl, dihydroisooxazolyl,dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl,dihydropyrazinyl, dihydropyrazolyl, dihydropyridinyl,dihydropyrimidinyl, dihydropyrrolyl, dihydroquinolinyl,dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl,dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl,methylenedioxybenzoyl, tetrahydrofuranyl, and tetrahydrothienyl, andN-oxides thereof.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

As used herein, “pharmaceutically acceptable salts” refer to derivativeswherein the parent compound is modified by making acid or base saltsthereof. Examples of pharmaceutically acceptable salts include, but arenot limited to, mineral or organic acid salts of basic residues such asamines; alkali or organic salts of acidic residues such as carboxylicacids; and the like. The pharmaceutically acceptable salts include theconventional non-toxic salts or the quaternary ammonium salts of theparent compound formed, for example, from non-toxic inorganic or organicacids. For example, such conventional non-toxic salts include thosederived from inorganic acids such as hydrochloric, hydrobromic,sulfuric, sulfamic, phosphoric, nitric and the like; and the saltsprepared from organic acids such as acetic, propionic, succinic,glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic,maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic,sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic,ethane disulfonic, oxalic, isethionic, and the like.

The pharmaceutically acceptable salts of the present invention can beprepared from the parent compound which contains a basic or acidicmoiety by conventional chemical methods. Generally, such salts can beprepared by reacting the free acid or base forms of these compounds witha stoichiometric amount of the appropriate base or acid in water or inan organic solvent, or in a mixture of the two; generally, nonaqueousmedia such as ether, ethyl acetate, ethanol, isopropanol, oracetonitrile are preferred. Suitable salts are found, e.g. inRemington's Pharmaceutical Sciences, 17 th ed., Mack Publishing Company,Easton, Pa., 1985, p. 1418.

Exemplifying the invention is the use of the compounds disclosed in theExamples and herein.

Specific compounds within the present invention include a compound whichselected from the group consisting of: the title compounds of theExamples;

and pharmaceutically acceptable salts thereof and individualdiastereomers thereof.

The subject compounds are useful in a method of modulating chemokinereceptor activity in a patient in need of such modulation comprising theadministration of an effective amount of the compound.

The present invention is directed to the use of the foregoing compoundsas modulators of chemokine receptor activity. In particular, thesecompounds are useful as modulators of the chemokine receptors, inparticular CCR-2.

The utility of the compounds in accordance with the present invention asmodulators of chemokine receptor activity may be demonstrated bymethodology known in the art, such as the assay for chemokine binding asdisclosed by Van Riper, et al., J. Exp. Med., 177, 851-856 (1993) whichmay be readily adapted for measurement of CCR-2 binding.

Receptor affinity in a CCR-2 binding assay was determined by measuringinhibition of ¹²⁵ I-MCP-1 to the endogenous CCR-2 receptor on variouscell types including monocytes, THP-1 cells, or after heterologousexpression of the cloned receptor in eukaryotic cells. The cells weresuspended in binding buffer (50 mM HEPES, pH 7.2, 5 mM MgCl₂, 1 mMCaCl₂, and 0.50% BSA or 0.5% human serum) and added to test compound orDMSO and ¹²⁵ I-MCP-1 at room temperature for 1 h to allow binding. Thecells were then collected on GFB filters, washed with 25 mM HEPES buffercontaining 500 mM NaCl and cell bound ¹²⁵ I-MCP-1 was quantified.

In a chemotaxis assay chemotaxis was performed using T cell depletedPBMC isolated from venous whole or leukophoresed blood and purified byFicoll-Hypaque centrifugation followed by rosetting withneuraminidase-treated sheep erythrocytes. Once isolated, the cells werewashed with HBSS containing 0.1 mg/ml BSA and suspended at 1×10⁷cells/ml. Cells were fluorescently labeled in the dark with 2 μMCalcien-AM (Molecular Probes), for 30 min at 37 ° C. Labeled cells werewashed twice and suspended at 5×10⁶ cells/ml in RPMI 1640 withL-glutamine (without phenol red) containing 0.1 mg/ml BSA. MCP-1(Peprotech) at 10 ng/ml diluted in same medium or medium alone wereadded to the bottom wells (27 μl). Monocytes (150,000 cells) were addedto the topside of the filter (30 μl) following a 15 min preincubationwith DMSO or with various concentrations of test compound. An equalconcentration of test compound or DMSO was added to the bottom well toprevent dilution by diffusion. Following a 60 min incubation at 37 ° C.,5% CO₂, the filter was removed and the topside was washed with HBSScontaining 0.1 mg/ml BSA to remove cells that had not migrated into thefilter. Spontaneous migration (chemokinesis) was determined in theabsence of chemoattractant.

In particular, the compounds of the following examples had activity inbinding to the CCR-2 receptor in the aforementioned assays, generallywith an IC₅₀ of less than about 1 μM. Such a result is indicative of theintrinsic activity of the compounds in use as modulators of chemokinereceptor activity.

Mammalian chemokine receptors provide a target for interfering with orpromoting eosinophil and/or leukocyte function in a mammal, such as ahuman. Compounds which inhibit or promote chemokine receptor function,are particularly useful for modulating eosinophil and/or leukocytefunction for therapeutic purposes. Accordingly, compounds which inhibitor promote chemokine receptor function would be useful in treating,preventing, ameliorating, controlling or reducing the risk of a widevariety of inflammatory and immunoregulatory disorders and diseases,allergic diseases, atopic conditions including allergic rhinitis,dermatitis, conjunctivitis, and asthma, as well as autoimmunepathologies such as rheumatoid arthritis and atherosclerosis, andfurther, chronic obstructive pulmonary disease, and multiple sclerosis.

For example, an instant compound which inhibits one or more functions ofa mammalian chemokine receptor (e.g., a human chemokine receptor) may beadministered to inhibit (i.e., reduce or prevent) inflammation. As aresult, one or more inflammatory processes, such as leukocyteemigration, chemotaxis, exocytosis (e.g., of enzymes, histamine) orinflammatory mediator release, is inhibited.

In addition to primates, such as humans, a variety of other mammals canbe treated according to the method of the present invention. Forinstance, mammals including, but not limited to, cows, sheep, goats,horses, dogs, cats, guinea pigs, rats or other bovine, ovine, equine,canine, feline, rodent or murine species can be treated. However, themethod can also be practiced in other species, such as avian species(e.g., chickens).

Diseases and conditions associated with inflammation and infection canbe treated using the compounds of the present invention. In a certainembodiment, the disease or condition is one in which the actions ofleukocytes are to be inhibited or promoted, in order to modulate theinflammatory response.

Diseases or conditions of humans or other species which can be treatedwith inhibitors of chemokine receptor function, include, but are notlimited to: inflammatory or allergic diseases and conditions, includingrespiratory allergic diseases such as asthma, particularly bronchialasthma, allergic rhinitis, hypersensitivity lung diseases,hypersensitivity pneumonitis, eosinophilic pneumonias (e.g., Loeffler'ssyndrome, chronic eosinophilic pneumonia), delayed-typehypersensitivity, interstitial lung diseases (ILD) (e.g., idiopathicpulmonary fibrosis, or ILD associated with rheumatoid arthritis,systemic lupus erythematosus, ankylosing spondylitis, systemicsclerosis, Sjogren's syndrome, polymyositis or dermatomyositis);systemic anaphylaxis or hypersensitivity responses, drug allergies(e.g., to penicillin, cephalosporins), insect sting allergies;autoimmune diseases, such as rheumatoid arthritis, psoriatic arthritis,multiple sclerosis, systemic lupus erythematosus, myasthenia gravis,juvenile onset diabetes; glomerulonephritis, autoimmune thyroiditis,Behcet's disease; graft rejection (e.g., in transplantation), includingallograft rejection or graft-versus-host disease; inflammatory boweldiseases, such as Crohn's disease and ulcerative colitis;spondyloarthropathies; scleroderma; psoriasis (including T-cell mediatedpsoriasis) and inflammatory dermatoses such an dermatitis, eczema,atopic dermatitis, allergic contact dermatitis, urticaria; vasculitis(e.g., necrotizing, cutaneous, and hypersensitivity vasculitis);eosinphilic myositis, eosinophilic fasciitis; and cancers, includingcancers with leukocyte infiltration of the skin or organs and othercancers. Inhibitors of chemokine receptor function may also be useful inthe treatment and prevention of stroke (Hughes et al., Journal ofCerebral Blood Flow & Metabolism, 22:308-317, 2002, and Takami et al.,Journal of Cerebral Blood Flow & Metabolism, 22:780-784, 2002),neurodegenerative conditions including but not limited to Alzheimer'sdisease, amyotrophic lateral sclerosis (ALS) and Parkinson's disease,obesity, type II diabetes, neuropathic and inflammatory pain, andGuillain Barre syndrome. Other diseases or conditions in whichundesirable inflammatory responses are to be inhibited can be treated,including, but not limited to, reperfusion injury, atherosclerosis,certain hematologic malignancies, cytokine-induced toxicity (e.g.,septic shock, endotoxic shock), polymyositis, dermatomyositis andchronic obstructive pulmonary disease.

Diseases or conditions of humans or other species, which can be treatedwith modulators of chemokine receptor function, include or involve butare not limited to: immunosuppression, such as that in individuals withimmunodeficiency syndromes such as AIDS or other viral infections,individuals undergoing radiation therapy, chemotherapy, therapy forautoimmune disease or drug therapy (e.g., corticosteroid therapy), whichcauses immunosuppression; immunosuppression due to congenital deficiencyin receptor function or other causes; and infections diseases, such asparasitic diseases, including, but not limited to helminth infections,such as nematodes (round worms), (Trichuriasis, Enterobiasis,Ascariasis, Hookworm, Strongyloidiasis, Trichinosis, filariasis),trematodes (flukes) (Schistosomiasis, Clonorchiasis), cestodes (tapeworms) (Echinococcosis, Taeniasis saginata, Cysticercosis), visceralworms, visceral larva migraines (e.g., Toxocara), eosinophilicgastroenteritis (e.g., Anisaki sp., Phocanema sp.), and cutaneous larvamigraines (Ancylostona braziliense, Ancylostoma caninum).

In addition, treatment of the aforementioned inflammatory, allergic,infectious and autoimmune diseases can also be contemplated for agonistsof chemokine receptor function if one contemplates the delivery ofsufficient compound to cause the loss of receptor expression on cellsthrough the induction of chemokine receptor internalization or deliveryof compound in a manner that results in the misdirection of themigration of cells.

The compounds of the present invention are accordingly useful intreating, preventing, ameliorating, controlling or reducing the risk ofa wide variety of inflammatory and immunoregulatory disorders anddiseases, allergic conditions, atopic conditions, as well as autoimmunepathologies. In a specific embodiment, the present invention is directedto the use of the subject compounds for treating, preventing,ameliorating, controlling or reducing the risk of autoimmune diseases,such as rheumatoid arthritis, psoriatic arthritis and multiplesclerosis.

In another aspect, the instant invention may be used to evaluateputative specific agonists or antagonists of chemokine receptors,including CCR-2. Accordingly, the present invention is directed to theuse of these compounds in the preparation and execution of screeningassays for compounds that modulate the activity of chemokine receptors.For example, the compounds of this invention are useful for isolatingreceptor mutants, which are excellent screening tools for more potentcompounds. Furthermore, the compounds of this invention are useful inestablishing or determining the binding site of other compounds tochemokine receptors, e.g., by competitive inhibition. The compounds ofthe instant invention are also useful for the evaluation of putativespecific modulators of the chemokine receptors, including CCR-2. Asappreciated in the art, thorough evaluation of specific agonists andantagonists of the above chemokine receptors has been hampered by thelack of availability of non-peptidyl (metabolically resistant) compoundswith high binding affinity for these receptors. Thus the compounds ofthis invention are commercial products to be sold for these purposes.

The present invention is further directed to a method for themanufacture of a medicament for modulating chemokine receptor activityin humans and animals comprising combining a compound of the presentinvention with a pharmaceutical carrier or diluent.

The present invention is further directed to the use of the presentcompounds in treating, preventing, ameliorating, controlling or reducingthe risk of infection by a retrovirus, in particular, herpes virus orthe human immunodeficiency virus (HIV) and the treatment of, anddelaying of the onset of consequent pathological conditions such asAIDS. Treating AIDS or preventing or treating infection by HIV isdefined as including, but not limited to, treating a wide range ofstates of HIV infection: AIDS, ARC (AIDS related complex), bothsymptomatic and asymptomatic, and actual or potential exposure to HIV.For example, the compounds of this invention are useful in treatinginfection by HIV after suspected past exposure to HIV by, e.g., bloodtransfusion, organ transplant, exchange of body fluids, bites,accidental needle stick, or exposure to patient blood during surgery.

In a further aspect of the present invention, a subject compound may beused in a method of inhibiting the binding of a chemokine to a chemokinereceptor, such as CCR-2, of a target cell, which comprises contactingthe target cell with an amount of the compound which is effective atinhibiting the binding of the chemokine to the chemokine receptor.

The subject treated in the methods above is a mammal, for instance ahuman being, male or female, in whom modulation of chemokine receptoractivity is desired. “Modulation” as used herein is intended toencompass antagonism, agonism, partial antagonism, inverse agonismand/or partial agonism. In an aspect of the present invention,modulation refers to antagonism of chemokine receptor activity. The term“therapeutically effective amount” means the amount of the subjectcompound that will elicit the biological or medical response of atissue, system, animal or human that is being sought by the researcher,veterinarian, medical doctor or other clinician.

The term “composition” as used herein is intended to encompass a productcomprising the specified ingredients in the specified amounts, as wellas any product which results, directly or indirectly, from combinationof the specified ingredients in the specified amounts. By“pharmaceutically acceptable” it is meant the carrier, diluent orexcipient must be compatible with the other ingredients of theformulation and not deleterious to the recipient thereof.

The terms “administration of” and or “administering a” compound shouldbe understood to mean providing a compound of the invention to theindividual in need of treatment.

As used herein, the term “treatment” refers both to the treatment and tothe prevention or prophylactic therapy of the aforementioned conditions.

Combined therapy to modulate chemokine receptor activity for therebytreating, preventing, ameliorating, controlling or reducing the risk ofinflammatory and immunoregulatory disorders and diseases, includingasthma and allergic diseases, as well as autoimmune pathologies such asrheumatoid arthritis and multiple sclerosis, and those pathologies notedabove is illustrated by the combination of the compounds of thisinvention and other compounds which are known for such utilities.

For example, in treating, preventing, ameliorating, controlling orreducing the risk of inflammation, the present compounds may be used inconjunction with an antiinflammatory or analgesic agent such as anopiate agonist, a lipoxygenase inhibitor, such as an inhibitor of5-lipoxygenase, a cyclooxygenase inhibitor, such as a cyclooxygenase-2inhibitor, an interleukin inhibitor, such as an interleukin-1 inhibitor,an NMDA antagonist, an inhibitor of nitric oxide or an inhibitor of thesynthesis of nitric oxide, a non-steroidal antiinflammatory agent, or acytokine-suppressing antiinflammatory agent, for example with a compoundsuch as acetaminophen, aspirin, codeine, biological TNF sequestrants,fentanyl, ibuprofen, indomethacin, ketorolac, morphine, naproxen,phenacetin, piroxicam, a steroidal analgesic, sufentanyl, sunlindac,tenidap, and the like. Similarly, the instant compounds may beadministered with a pain reliever; a potentiator such as caffeine, anH2-antagonist, simethicone, aluminum or magnesium hydroxide; adecongestant such as phenylephrine, phenylpropanolamine,pseudoephedrine, oxymetazoline, ephinephrine, naphazoline,xylometazoline, propylhexedrine, or levo-desoxy-ephedrine; anantiitussive such as codeine, hydrocodone, caramiphen, carbetapentane,or dextramethorphan; a diuretic; and a sedating or non-sedatingantihistamine.

Likewise, compounds of the present invention may be used in combinationwith other drugs that are used in the treatment/prevention/suppressionor amelioration of the diseases or conditions for which compounds of thepresent invention are useful. Such other drugs may be administered, by aroute and in an amount commonly used therefore, contemporaneously orsequentially with a compound of the present invention. When a compoundof the present invention is used contemporaneously with one or moreother drugs, a pharmaceutical composition containing such other drugs inaddition to the compound of the present invention may be used.Accordingly, the pharmaceutical compositions of the present inventioninclude those that also contain one or more other active ingredients, inaddition to a compound of the present invention.

Examples of other active ingredients that may be combined with CCR2antagonists, such as the CCR2 antagonists compounds of the presentinvention, either administered separately or in the same pharmaceuticalcompositions, include, but are not limited to: (a) VLA-4 antagonistssuch as those described in U.S. Pat. No. 5,510,332, WO95/15973,WO96/01644, WO96/06108, WO96/20216, WO96/22966, WO96/31206, WO96/40781,WO97/03094, WO97/02289, WO 98/42656, WO98/53814, WO98/53817, WO98/53818,WO98/54207, and WO98/58902; (b) steroids such as beclomethasone,methylprednisolone, betamethasone, prednisone, dexamethasone, andhydrocortisone; (c) immunosuppressants such as cyclosporin, tacrolimus,rapamycin, EDG receptor agonists including FTY-720, and other FK-506type immunosuppressants; (d) antihistamines (H1-histamine antagonists)such as bromopheniramine, chlorpheniramine, dexchlorpheniramine,triprolidine, clemastine, diphenhydramine, diphenylpyraline,tripelennamine, hydroxyzine, methdilazine, promethazine, trimeprazine,azatadine, cyproheptadine, antazoline, pheniramine pyrilamine,astemizole, terfenadine, loratadine, desloratadine, cetirizine,fexofenadine, descarboethoxyloratadine, and the like; (e) non-steroidalanti-asthmatics such as β2-agonists (terbutaline, metaproterenol,fenoterol, isoetharine, albuterol, bitolterol, and pirbuterol),theophylline, cromolyn sodium, atropine, ipratropium bromide,leukotriene antagonists (zafirlukast, montelukast, pranlukast,iralukast, pobilukast, SKB-106,203), leukotriene biosynthesis inhibitors(zileuton, BAY-1005); (f) non-steroidal antiinflammatory agents (NSAIDs)such as propionic acid derivatives (alminoprofen, benoxaprofen, bucloxicacid, carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen,ibuprofen, indoprofen, ketoprofen, miroprofen, naproxen, oxaprozin,pirprofen, pranoprofen, suprofen, tiaprofenic acid, and tioxaprofen),acetic acid derivatives (indomethacin, acemetacin, alclofenac, clidanac,diclofenac, fenclofenac, fenclozic acid, fentiazac, furofenac, ibufenac,isoxepac, oxpinac, sulindac, tiopinac, tolmetin, zidometacin, andzomepirac), fenamic acid derivatives (flufenamic acid, meclofenamicacid, mefenamic acid, niflumic acid and tolfenamic acid),biphenylcarboxylic acid derivatives (diflunisal and flufenisal), oxicams(isoxicam, piroxicam, sudoxicam and tenoxican), salicylates (acetylsalicylic acid, sulfasalazine) and the pyrazolones (apazone,bezpiperylon, feprazone, mofebutazone, oxyphenbutazone, phenylbutazone);(g) cyclooxygenase-2 (COX-2) inhibitors; (h) inhibitors ofphosphodiesterase type IV (PDE-IV); (i) other antagonists of thechemokine receptors, especially CCR-1, CCR-2, CCR-3, CXCR-3, CXCR-4 andCCR-5; (j) cholesterol lowering agents such as HMG-CoA reductaseinhibitors (lovastatin, simvastatin and pravastatin, fluvastatin,atorvastatin, rosuvastatin, and other statins), sequestrants(cholestyramine and colestipol), cholesterol absorption inhibitors(ezetimibe), nicotinic acid, fenofibric acid derivatives (gemfibrozil,clofibrat, fenofibrate and benzafibrate), and probucol; (k)anti-diabetic agents such as insulin, sulfonylureas, biguanides(metformin), α-glucosidase inhibitors (acarbose) and glitazones(troglitazone and pioglitazone); (1) preparations of interferon beta(interferon beta-1α, interferon beta-1β); (m) preparations of glatirameracetate; (n) preparations of CTLA4 Ig; (o) preparations ofhydroxychloroquine, (p) Copaxone® and (q) other compounds such as5-aminosalicylic acid and prodrugs thereof, antimetabolites such asazathioprine, 6-mercaptopurine and methotrexate, leflunomide,teriflunomide, and cytotoxic and other cancer chemotherapeutic agents.

The weight ratio of the compound of the present invention to the secondactive ingredient may be varied and will depend upon the effective doseof each ingredient. Generally, an effective dose of each will be used.Thus, for example, when a compound of the present invention is combinedwith an NSAID the weight ratio of the compound of the present inventionto the NSAID will generally range from about 1000:1 to about 1:1000, orfrom about 200:1 to about 1:200. Combinations of a compound of thepresent invention and other active ingredients will generally also bewithin the aforementioned range, but in each case, an effective dose ofeach active ingredient should be used.

In such combinations the compound of the present invention and otheractive agents may be administered separately or in conjunction. Inaddition, the administration of one element may be prior to, concurrentto, or subsequent to the administration of other agent(s).

The compounds of the present invention may be administered by oral,parenteral (e.g., intramuscular, intraperitoneal, intravenous, ICV,intracisternal injection or infusion, subcutaneous injection, orimplant), by inhalation spray, nasal, vaginal, rectal, sublingual, ortopical routes of administration and may be formulated, alone ortogether, in suitable dosage unit formulations containing conventionalnon-toxic pharmaceutically acceptable carriers, adjuvants and vehiclesappropriate for each route of administration. In addition to thetreatment of warm-blooded animals such as mice, rats, horses, cattle,sheep, dogs, cats, monkeys, etc., the compounds of the invention areeffective for use in humans.

The pharmaceutical compositions for the administration of the compoundsof this invention may conveniently be presented in dosage unit form andmay be prepared by any of the methods well known in the art of pharmacy.All methods include the step of bringing the active ingredient intoassociation with the carrier which constitutes one or more accessoryingredients. In general, the pharmaceutical compositions are prepared byuniformly and intimately bringing the active ingredient into associationwith a liquid carrier or a finely divided solid carrier or both, andthen, if necessary, shaping the product into the desired formulation. Inthe pharmaceutical composition the active object compound is included inan amount sufficient to produce the desired effect upon the process orcondition of diseases. As used herein, the term “composition” isintended to encompass a product comprising the specified ingredients inthe specified amounts, as well as any product which results, directly orindirectly, from combination of the specified ingredients in thespecified amounts.

The pharmaceutical compositions containing the active ingredient may bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsions, hard or soft capsules, or syrups or elixirs. Compositionsintended for oral use may be prepared according to any method known tothe art for the manufacture of pharmaceutical compositions and suchcompositions may contain one or more agents selected from the groupconsisting of sweetening agents, flavoring agents, coloring agents andpreserving agents in order to provide pharmaceutically elegant andpalatable preparations. Tablets contain the active ingredient inadmixture with non-toxic pharmaceutically acceptable excipients whichare suitable for the manufacture of tablets. These excipients may be forexample, inert diluents, such as calcium carbonate, sodium carbonate,lactose, calcium phosphate or sodium phosphate; granulating anddisintegrating agents, for example, corn starch, or alginic acid;binding agents, for example starch, gelatin or acacia, and lubricatingagents, for example magnesium stearate, stearic acid or talc. Thetablets may be uncoated or they may be coated by known techniques todelay disintegration and absorption in the gastrointestinal tract andthereby provide a sustained action over a longer period. For example, atime delay material such as glyceryl monostearate or glyceryl distearatemay be employed. They may also be coated by the techniques described inthe U.S. Pat. Nos. 4,256,108; 4,166,452 ; and 4,265,874 to form osmotictherapeutic tablets for control release.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxy-propylmethylcellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethylene-oxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl, p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, forexample olive oil or arachis oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative and flavoring and coloringagents.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally-acceptable diluent orsolvent, for example as a solution in 1,3-butane diol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose any bland fixed oil may be employedincluding synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid find use in the preparation of injectables.

The compounds of the present invention may also be administered in theform of suppositories for rectal administration of the drug. Thesecompositions can be prepared by mixing the drug with a suitablenon-irritating excipient which is solid at ordinary temperatures butliquid at the rectal temperature and will therefore melt in the rectumto release the drug. Such materials are cocoa butter and polyethyleneglycols.

For topical use, creams, ointments, jellies, solutions or suspensions,etc., containing the compounds of the present invention are employed.(For purposes of this application, topical application shall includemouthwashes and gargles.).

The pharmaceutical composition and method of the present invention mayfurther comprise other therapeutically active compounds as noted hereinwhich are usually applied in the treatment of the above mentionedpathological conditions.

In treating, preventing, ameliorating, controlling or reducing the riskof conditions which require chemokine receptor modulation an appropriatedosage level will generally be about 0.0001 to 500 mg per kg patientbody weight per day which can be administered in single or multipledoses. In certain embodiments the dosage level will be about 0.0005 toabout 400 mg/kg per day; or from about 0.005 to about 300 mg/kg per day;or from about 0.01 to about 250 mg/kg per day, or from about 0.05 toabout 100 mg/kg per day, or from about 0.5 to about 50 mg/kg per day.Within this range the dosage may be 0.0001 to 0.005, 0.005 to 0.05, 0.05to 0.5, 0.5 to 5 or 5 to 50 mg/kg per day. For oral administration, thecompositions may be provided in the form of tablets containing 0.01 to1000 milligrams of the active ingredient, or 0.1 to 500, 1.0 to 400, or2.0 to 300, or 3.0 to 200, particularly 0.01, 0.05, 0.1, 1, 4, 5, 10,15, 20, 25, 30, 50, 75, 100, 125, 150, 175, 200, 250, 300, 400, 500,600, 750, 800, 900, and 1000 milligrams of the active ingredient for thesymptomatic adjustment of the dosage to the patient to be treated. Thecompounds may be administered on a regimen of 1 to 4 times per day, oronce or twice per day.

It will be understood, however, that the specific dose level andfrequency of dosage for any particular patient may be varied and willdepend upon a variety of factors including the activity of the specificcompound employed, the metabolic stability and length of action of thatcompound, the age, body weight, general health, sex, diet, mode and timeof administration, rate of excretion, drug combination, the severity ofthe particular condition, and the host undergoing therapy.

Several methods for preparing the compounds of this invention areillustrated in the following Schemes and Examples. Starting materialsare made by known procedures or as illustrated.

Several methods for preparing the compounds of this invention areillustrated in the following Schemes and Examples. Starting materialsare either commercially available or made by known procedures in theliterature or as illustrated. The present invention further providesprocesses for the preparation of compounds of the formula I as definedabove, which comprises many different sequences of assembling compoundsof formula (II), formula (III), and formula (IV), or compounds offormula (V), formula (VI), and formula (IV), or compounds of formula(VII) and formula (IV).

wherein R¹, R³, R⁵, R⁸, R¹⁰, and X are defined as in formula I, andR^(10a) represents either a hydrogen or an alkyl group such as methyl,ethyl, t-butyl, or benzyl which serves as a protecting group, R⁷represent either hydrogen or an amine protecting group (Greene, T; Wuts,P. G. M. Protective Groups in Organic Synthesis, John Wiley & Sons,Inc., New York, N.Y. 1991) such as Boc or trifluoroacetate. The bondbetween the two carbon atoms where a dashed line is shown in formula IIIand in formula VII represent either a single or double bond as definedin formula I.

One general way of constructing target compounds I utilizingIntermediates of the formulas II, III, and IV is illustrated inScheme 1. Coupling of the acid IIIa and the amine IV under standardamide bond formation reaction conditions such as PyBrop in the presenceof a base such as N,N-diisopropylethylamine and a catalyst such as DMAPgives the intermediate 1-1. Removal of the Boc protecting group yieldsthe amine 1-2. Reductive alkylation of 1-2 with ketones II in thepresence of a borohydride such as sodium triacetoxyborohydride or sodiumcyanoborohydride then provides the compound of formula Ia. Note thatwhen R⁸ or R¹⁰ are other than hydrogen, a mixture of diastereomers(Eliel, E. E., Wilen, S. H., Stereochemistry of Organic Compounds, JohnWiley & Sons, Inc., New York) results from the reductive amination step.These can be separated into their components by chromatography usingnormal phase, reverse phase or chiral columns, depending on the natureof the separation. Compound Ia can be further elaborated to the compoundof the formula I by reductive alkylation with an aldehyde or byalkylation with, for example, an alkyl halide.

In some cases Intermediate 1-1 may require modification prior toelaboration to 1-2. For example (see below) oxidation of the5-azatetrahydroisoquinoline moiety (where R³ in 1-1 is nothing) to itsN-oxide (where R³ ═O) may be conveniently performed at this stage togive 1-1a. This can be accomplished with a variety of oxidants includingmCPBA. Compound 1-1a can be carried on in the same fashion as 1-1 inScheme 1 to give Ia.

An alternative sequence of construction involving fragments of theformulas II, III, and W is depicted in Scheme 1A. Amine IIIb isreductively alkylated with ketone II in the presence of a borohydridesuch as sodium triacetoxyborohydride or sodium cyanoborohydride to givesecondary amine 1-3. Protection of the amine group can be accomplishedusing any of a number of protecting groups, including thetrifluoroacetamide group (R¹² ═COCF₃), which can be installed bytreatment with trifluoroacetic anhydride in the presence of a base suchas triethylamine. The ester functionality of the resulting compound 1-4is then cleaved using conditions that are dependent upon the nature ofR^(10a). For example, a benzyl ester is cleaved by hydrogenolysis usinga catalyst such as Pd on carbon to give the fragment of the formula VII.Coupling of the acid VII and the amine IV under standard amide bondformation reaction conditions such as PyBrop in the presence of a basesuch as N,N-diisopropylethylamine and a catalyst such as DMAP gives theintermediate 1-5. Alternatively, the acid VII can be converted to itscorresponding acid chloride and then treated with amine IV in thepresence of a base such as triethylamine to give 1-5. Removal of theprotecting group (R¹²) to give compound Ia can be achieved in variousways depending upon the nature of the protecting group. For example thetrifluoroacetate group can be removed by treatment with excess sodiumborohydride, or by treatment with a base such as lithium hydroxide.

Alternatively, Intermediate 1-3 from Scheme 1A can be more directlyaccessed as shown in Scheme 1B. In this case amine IIIc is reductivelyalkylated with ketone II in the presence of a borohydride such as sodiumtriacetoxyborohydride or sodium cyanoborohydride to give secondary amine1-3a. Treatment with a base such as LDA then generates the enolate of1-3a which can be alkylated with a variety of electrophiles includingbut not limited to alkyl halides, aldehydes, ketones. The resultingcompound 1-3 can be carried on to compounds of the formula I or Ia,using the same steps as outlined in Scheme 1A.

In addition to assembly according to Schemes 1, and 1A-1B, compounds ofthe formula I can be prepared using Intermediates of the formula IV, Vand VII (Scheme 2). According to this protocol, known keto acid VIa issimultaneously converted to dimethyl acetal-ester VIb usingtrimethylorthoformate, methanol and an acid catalyst such as toluenesulfonic acid. Alkylation of 2-1 can be carried out with a base such asLDA and an electrophile such as an alkyl halide to give 2-2. Hydrolysisof the methyl ester and removal of the dimethyl acetal protecting groupcan be accomplished by treatment with a base such as NaOH, followed byan acid such as HCl. The resulting acid VIb can be coupled to amine IVusing various conditions. For example acid VIb can be converted to itscorresponding acid chloride with oxalyl chloride and catalytic DMF, thentreated with amine IV. The amide 2-3 can be resolved using chiral HPLCto give a single enantiomer 2-3a. Reductive amination of 2-3a with amineV using, for example, NaB(OAc)₃H gives target compound Ia, which can, ifappropriate, be further modified to compounds I as shown in Scheme 1.Note that the compound Ia formed in the above mentioned transformationwas obtained initially as a mixture of 1,3-cis- and1,3-trans-diastereoisomers. These could be separated into the respectivesingle diastereoisomers in various ways, including by preparative TLC,column chromatography, and chiral HPLC to provide the preferred1,3-cis-isomer Ia shown.

An alternate route to homochiral 2-3a involves oxidation of aminoacidIIId as shown in Scheme 2A. This transformation can be accomplishedusing NBS as the oxidant. The resulting keto acid VIc is obtained as asingle enantiomer in this way, which can then be carried on toIntermediate 2-3a, and ultimately to compound Ia and I, as shown inScheme 2.

The cyclopentane core fragment III can be prepared in a number of ways.One of those is depicted in Scheme 3, 3a, and 3b. According to Scheme 3,the commercially available homochiral lactam 3-1 is hydrogenated and thesaturated 3-2 is treated with di-tert-butyl dicarbonate in the presenceof a suitable catalyst, e.g. N,N-dimethylamino pyridine. A basecatalyzed cleavage of the amide bond in the presence of a suitablealcohol R^(10a)—OH then provides the respective ester IIIe. TheBOC-protecting group is removed, preferably with an acid such as HCl ina aprotic solvent, such as dioxane, to yield the amine IIIf in the formof a salt. When this amine is mixed with benzophenone imine, therespective Schiff base IIIg is formed, which can be obtained in pureform by simple filtration to remove ammonium chloride.

The enolate formed from ester IIIg with a strong base, such as LDA canbe reacted with alkyl halides R¹—X, as well as aldehydes R^(1a)CHO orketones R^(1a)R^(2a) CO to obtain intermediates IIIh, 3-4 and IIIi, 3-5respectively, Scheme 3A. These reactions produce a mixture of therespective cis-(IIIh and IIIi) and trans-(3-4 and 3-5) diastereoisomers,which can be separated by a suitable chromatography. In most cases,normal phase flash chromatography on deactivated silica gel can beapplied with success.

The desired cis diastereoisomers IIIh and IIIi are then treated with anacid such as HCl to aid hydrolysis of the imine group and the resultingamino group IIIj can be suitably protected e.g. in a form of atert-butoxycarbonyl amide (Scheme 3B). The ester group present inintermediates IIIk can then be cleaved to give acid IIIl. The appliedprocedure depends on the nature of the ester: e.g. a benzyl ester can becleaved by hydrogenolysis, a tert-butyl ester under acidic conditionsand a alkyl ester can be hydrolyzed under either acidic or basicconditions.

Note that Compound IIIl can be used in place of IIIa in Scheme 1, IIIjcan be used in place of IIIb in Scheme 1A, and IIIf can be used in placeof IIIc in Scheme 1B (the only differences being that the cyclopentanerings are defined as being fully saturated). An alternative way ofpreparing compounds of the type III is shown in Scheme 3C. According tothis route, commercially available m is converted to ester IIIn using anappropriate alcohol such as methyl or benzyl alcohol in the presence ofan acid catalyst. Protection of the amine in IIIn by treatment withBoc₂O results in IIIo. Alkylation using a base such as lithiumhexamethyldisilazide (LiHMDS) and an electrophile such as an alkylhalide gives IIIp, where the major diastereomer obtained is normally thecis-1,3-isomer. Separation of the cis/trans isomers can be carried outat this point or after the following step using column chromatography.If desired, hydrogenation using a catalysts such as Pd/C gives IIIq. IfR¹⁰ is benzyl hydrogenation of IIIp would directly furnish IIIr.Otherwise, IIIq can be hydrolyzed using various conditions such astreatment with NaOH to give IIIr. If desired IIIr can be treated withHCl or TFA to give IIId (used in Scheme 2a).

The 5-aza-tetrahydroisoquinoline fragment IV can be prepared in severalways, including in accordance to the literature methods of MarCoux, J-F.et al. (J. Chem. Lett., 2000, 2 (15), 2339-2341). Alternatively,fragment IV can be prepared as outlined in Scheme 4. Compound 4-1,normally obtained from commercial sources, is brominated (Br₂, AcOH) togive 4-2. Metal halogen exchange (NaH, t-butyl lithium) followed bytreatment with DMF provides aldehyde 4-3. Conversion of the aldehydegroup to a nitrile can be achieved with sodium formate, hydroxylaminehydrochloride and formic acid. The resulting nitrile 4-4 can be treatedwith phosphorous oxychloride to give 2-chloropyridine 4-5. Displacementof the chloro group can be achieved with the sodium salt of adialkylmalonate. Reduction of the nitrile group of 4-6 with hydrogen andRaney Ni catalyst is accompanied by cyclization to afford compound 4-7.Decarboxylation can be achieved in a variety of ways depending on theester. In the case represented in Scheme 4, the t-butyl ester wasdecarboxylated with TFA to give 4-8. Reduction (BH₃), followed byprotection of the resulting amine using Boc₂O, gives 4-9, which can beconveniently purified. Removal of the Boc protecting group to give IVacan be achieved in various ways, including by treatment with anhydrousHCl in dioxane or some other solvent.

Compounds of the type IV could also be prepared according to Scheme 4A.Commercially available 4-10 can be methylated with methyl iodide in thepresence of a base such as K₂CO₃ to give 4-11. Cycloaddition with aprotected piperidinone in the presence of NH₃ in methanol furnishes5-azatetrahydroiso-quinoline 4-12 (R²² can be various protecting groupssuch as benzyl or benzoyl). Hydrogenation of the nitro group of compound4-12 with hydrogen and a catalyst such as Pd/C gives 4-13. Diazoniumsalt formation followed by warming with sulfuric acid provides5-aza-7-hydroxytetrahydroisoquinoline 4-14. Removal of the protectinggroup R²² is achieved in different ways depending upon the nature ofR²². If R²² is benzyl, hydrogenation in the presence of HCl and acatalyst such as Pd/C can be applied. If R²² is benzoyl, hydrolysis canbe achieved by heating in concentrated HCl solution. Installation of aBoc protecting group on to 4-15 can be easily achieved with Boc₂O togive 4-16. Various R²³ can then be incorporated generating ethers (seeScheme 4B). The Boc protecting group on the resulting compounds 4-17 canfinally be removed with HCl or TFA to give IVb. Alternatively, Compound4-14 itself can be converted to ethers (according to Scheme 4B). Theresulting ether 4-18 can be converted to compound IVb by removal of R²²as described above.

The 5-aza-7-hydroxytetrahydroisoquinolines 4-14 and 4-16 in Scheme 4Acan be converted to various ethers (see Scheme 4B). Alkyl ethers can begenerated from an alkyl halide and a base (such as K₂CO₃, NaOH, or NaH)giving compounds 4-19 and 4-22. A trifluoromethyl ether can be preparedby initial methyl xanthate formation (NaH, CS₂; MeI), followed bysequential treatment with 1,3-dibromo-5,5-dimethylhydantoin (or NBS) andHF/pyridine solution giving 4-20. Aryl ethers can be prepared by anumber of methods, including reaction of arylboronic acids in thepresence of copper (II) acetate and triethylamine, to give compounds4-21.

Compounds 1V where R⁵ is a halide (IVc) can be prepared according toScheme 4C. Compound 4-13 can be converted to the halide 4-22 accordingto classical procedures via the diazonium salt. Alternatively the knowncycloaddition reaction to a suitably protected piperidinone can beapplied. Removal of the protecting group R²² can be achieved asdescribed previously.

After incorporation into advanced intermediates, fragments IVc can befurther modified so as to prepared 7-aryl-5-azotetrahydroisoquinolinecontaining analogs (Scheme 4D). This can be accomplished by coupling ofthe 5-aza-7-halotetrahydroisoquinoline intermediates 1-5a to arylboronic acids (or aryl stannanes), mediated by transition metalcatalysts such as Pd(OAc)₂.

Compounds of the type represented by fragment II were often commerciallyavailable, but sometimes required preparation. For example, compoundsIIa (Scheme 5) where X is either CH₂, S, O, or NP (P=protecting group)are commercially available. Compounds can be easily modified to IIb,having R⁸ groups, where R⁸ is an alkyl group, by deprotonation with abase such as LDA and alkylation with an alkyl halide (for a publishedprocedure involving tetrahydropyran-4-one see J. Am. Chem. Soc., 1991,113, 2079-2089). Compounds IIb can be incorporated into final targetcompounds as shown in the preceding Schemes. Sometimes furthermodification of R⁸ can be carried out. For example, If R⁸ is an allylgroup, oxidative cleavage (O₃; DMS or another method) gives thedicarbonyl compound IIc, which can undergo a double reductive aminationcyclization sequence as shown in Scheme 5A.

A synthesis of ketones IId where R⁸ is an alkoxy group is detailed inScheme 5B. According to this, commercially available5,6-dihydro-4-methoxy-2H-pyran (5-1) is treated with m-chloroperbenzoicacid in methanol to affect direct conversion to 5-2. An alkylation ofthe secondary alcohol with an appropriate alkyl halide R²⁵X in apresence of a base such as sodium hydride affords the ether 5-3.Deprotection of the acetal under acidic conditions affords the desiredketones IId. In this manner, a number of 3-alkoxyderivatives can besynthesized. Alternatively, 5-2 can be itself deprotected to give3-hydroxy-tetrahydropyran-4-one IIe. Further details, as well asexamples are described in the Experimental section.

Alternatively, Intermediates 5-2 can be prepared in an asymmetricfashion according to Scheme 5C. Enol benzoate 54 can be prepared fromketone IIf by trapping the enolate generated upon treatment with a basesuch as KHMDS with benzoic anhydride. Asymmetric oxidation can beaccomplished according to the conditions described by Yian Shi, et al.(J. Org. Chem., 2001, 66, 1818-1826) to give 5-6 as predominantly asingle isomer. Ring opening of the epoxide and generation of thedimethyl acetal occurs in one pot by treatment with an acid such as CSAin methanol to give 5-2a. Either enantiomer of 5-2 could be obtained byappropriate choice of the sugar catalyst 5-5.

Compounds of the type V can often be obtained from commercial sources.Alternatively, compounds V can be prepared from Intermediates IIaccording to Scheme 6. Reductive amination of II with an amine such asaminodiphenylmethane using a hydride source such as NaB(OAc)₃H orNaBH₃CN gives 6-1. Compounds 6-1, if warranted, can be resolved intoindividual isomers by various means, including crystallization with achiral acid and chiral HPLC. Removal of the diphenylmethyl protectinggroup with hydrogen in the presence of a catalyst (such as Pd/C) affordssubunit V.

In some cases the order of carrying out the foregoing reaction schemesmay be varied to facilitate the reaction or to avoid unwanted reactionproducts. The following examples are provided for the purpose of furtherillustration only and are not intended to be limitations on thedisclosed invention.

Concentration of solutions was generally carried out on a rotaryevaporator under reduced pressure. Flash chromatography was carried outon silica gel (230-400 mesh). MPLC refers to medium pressure liquidchromatography and was carried out on a silica gel stationary phaseunless otherwise noted. NMR spectra were obtained in CDCl₃ solutionunless otherwise noted. Coupling constants (J) are in hertz (Hz).Abbreviations: diethyl ether (ether), triethylamine (TEA),N,N-diisopropylethylamine (DIEA) saturated aqueous (sat'd), roomtemperature (rt), hour(s) (h), minute(s) (min).

The following are representative procedures for the preparation of thecompounds used in the following Examples or which can be substituted forthe compounds used in the following Examples which may not becommercially available.

In some cases the order of carrying out the foregoing reaction schemesmay be varied to facilitate the reaction or to avoid unwanted reactionproducts. The following examples are provided for the purpose of furtherillustration only and are not intended to be limitations on thedisclosed invention.

Concentration of solutions was generally carried out on a rotaryevaporator under reduced pressure. Flash chromatography was carried outon silica gel (230-400 mesh). NMR spectra were obtained in CDCl₃solution unless otherwise noted. Coupling constants (J) are in hertz(Hz). Abbreviations: diethyl ether (ether), triethylamine (TEA),N,N-diisopropylethylamine (DIEA) saturated aqueous (sat'd), roomtemperature (rt), hour(s) (h), minute(s) (min).

The following are representative Procedures for the preparation of thecompounds used in the following Examples or which can be substituted forthe compounds used in the following Examples which may not becommercially available.

Procedures for the Preparation of Non-carbamate Piperidines.

Intermediate 1 was prepared according to the procedure described in J.Am. Chem. Soc., 1991, 113, 2079-2089.

To a solution of terahydro-4H-pyran-4-one (5.0 g, 50 mmol) andhexamethylphosphoramide (8.70 mL) in tetrahydrofuran (150 mL) was addedslowly a solution of lithium diisopropylamide (31.25 mL, 2 M solution)in 125 mL of tetrahydrofuran at −78 ° C. The reaction mixture wasstirred for 5 min and then ethyl iodide was added (16.0 mL, 200 mmol).The mixture was gradually warmed to 0 ° C. over 2 h. The reactionmixture was quenched with a saturated solution of NH₄Cl and thenextracted with ether (4×100 mL). The ether layer was washed with brine,dried (anhydrous magnesium sulfate), concentrated, and purified by flashcolumn chromatography eluting with hexanes/ethyl acetate (4:1) to giveIntermediate 2 (1.20 g, 20%).

To a mixture of 5,6-dihydro-4-methoxy-2H-pyran (10.0 g, 87.5 mmol) inmethanol (200 mL) at 0 ° C. was added dropwise a solution of3-chloroperoxy-benzoic acid (30.2 g, 175 mmol) in methanol (50 mL) viaan addition funnel. The resulting solution was stirred for 5 h allowingit to warm to room temperature. The methanol was removed under reducedpressure affording a white solid. The material was dissolved in 500 mLof dichloromethane and cooled to 0° C. To the mixture, while stirringvigorously, was added in portions an excess of solid calcium hydroxide(50-60 g). After stirring an additional 30 min, the mixture was filteredthrough a plug of celite and the filtrate was evaporated under reducedpressure to afford 11.62 g (82%) of the desired product as a clear oil.¹H NMR (500 MHz, CDCl₃) δ 3.88-3.80 (m, 2H), 3.73-3.68 (m, 2H),3.54-3.48 (m, 1H), 3.28 (s, 3H), 3.27 (s, 3H), 2.00-1.93 (m, 1H),1.82-1.77 (m, 1H).

To a cooled (0° C.) solution of the product from Step A, Intermediate 3(9.40 g, 58.0 mmol) in tetrahydrofuran (200 mL), under nitrogen, wasslowly added NaH (2.32 g, 58.0 mmol) and the resulting slurry wasstirred for 1 h at 0° C. Iodomethane (7.22 mL, 116 mmol) was then addedvia syringe to the slurry and the resulting mixture was stirredovernight allowing it to warm to room temperature. The reaction wasquenched with a saturated solution of ammonium chloride (200 mL) and theorganic layer was then removed using a separatory funnel. The aqueouslayer was extracted with ether (3×150 mL) and all the organics werecombined, dried over anhydrous sodium sulfate, filtered, and evaporatedin vacuo. Purification was accomplished by flash column using a stepwisegradient eluant of 10-60% ether/hexanes to afford 8.46 g (83%) of thedesired product as a clear oil. ¹H NMR (500 MHz, CDCl₃) δ 3.98 (dd,J=2.5, 12.4 Hz, 1H), 3.77 (ddd, J=3.5, 7.1, 10.8 Hz, 1H), 3.57 (dd,J=1.4, 12.4 Hz, 1H), 3.50 (dd, J=2.5, 11.7 Hz, 1H), 3.46 (s, 3H), 3.25(s, 3H), 3.22 (s, 3H), 3.22-3.20 (m, 1H), 1.96 (ddd, J=4.7, 11.8, 16.5Hz, 1H), 1.75 (br dd, J=1.7, 14.2 Hz, 1H).

A solution of the product from Step B, Intermediate 3 (3.0 g, 17.04mmol) in tetrahydrofuran/water (60 mL/10 mL) was treated withconcentrated hydrochloric acid (6 mL) and the resulting solution wasstirred at room temperature for 1 h. The mixture was concentrated invacuo to remove the tetrahydrofuran and the aqueous layer then extractedwith ether (6×50 mL). The organics were combined, dried over anhydroussodium sulfate, filtered, and evaporated under reduced pressure toafford intermediate 24 (1.75 g, 79%) as a clear oil. ¹H NMR (500 MHz,CDCl₃) δ 4.23 (ddd, J=1.2, 11.4, 12.4 Hz, 1H), 4.15-4.09 (m, 1H), 3.82(dd, J=5.95, 8.7 Hz, 1H), 3.74 (ddd, J=5.5, 8.5, 13.6 Hz, 1H), 3.56 (dd,J=8.8, 11.3 Hz, 1H), 3.50 (s, 3H), 2.61 (app dd, J=5.0, 8.9 Hz, 2H).

This intermediate was prepared in an analogous fashion to that ofIntermediate 3, except iodomethane was replaced with iodoethane.Purification by MPLC (gradient elution from 040% ethyl acetate/hexanes)afforded 683 mg (66%) of the final compound as a clear oil.

To a suspension of Na₂HPO₄ (24.85 g, 175.1 mmol) and5,6-dihydro-4-methoxy-2H-pyran (10.0 g, 87.5 mmol) in dichloromethane(200 mL) at 0° C. was added dropwise a solution of 3-chloroperoxybenzoicacid (30.2 g, 175 mmol) in dichloromethane (50 mL) via addition funnel.The resulting solution was stirred for 5 h allowing it to warm to roomtemperature. The reaction was quenched with water (200 mL) and theorganics were separated. The aqueous layer was extracted withdichloromethane (200 mL) and the organics combined, dried over anhydroussodium sulfate, filtered, and evaporated in vacuo to afford Intermediate5 (19.12 g, 86%) as a white solid.

To a mixture of 5.6-dihydro-4-methoxy-2H-pyran (0.5 g, 4 mmol) inacetonitrile/water (15 mL, 1:1) at room temperature was added1-(chloromethyl)-4-fluoro-1,4-diazoniabicyclo[2.2.2.]octanebis(tetrafluoroborate) (1.5 g, 4.4 mmol, SELECTFLUOR™) in one lot andthe resulting reaction mixture was stirred at room temperature untilcompletion. Solid NaCl was then added and the reaction mixture was thenextracted with ether (4×50 mL). The ether layer was dried (anhydrousmagnesium sulfate) and concentrated to yield 0.34 g (65%) of the titlecompound that required no further purification. ¹H NMR (500 MHz, CDCl₃):d 4.95 (m, 1H), 4.44.21 (m, 2H), 3.72-3.65 (m, 2H), 2.75 (m, 2H).

A mixture of tetrahydro-4H-pyran-4-one (10.0 g, 100 mmol) andpyrrolidine (11 g, 150 mmol) was stirred at room temperature for 1 h.The excess pyrrolidine was removed in vacuo and the residue was driedovernight under high vacuum. The enamine was obtained as a yellow liquid(14.7 g) which was used in the next step without further purification.

The enamine, prepared in Step A, Intermediate 7 (1.54 g, 10 mmol) and4-N,N-dimethylpyridine (1.22 g) were treated with N,N-dimethylformamide(25 mL). The mixture was cooled to 0° C. and solid5-(trifluoromethyl)dibenzothiophenium trifluoromethanesulfonate (4.0 g,10 mmol) was added. The resulting mixture was stirred at 0° C. for 1 h,then quenched with 30 mL of concentrated aqueous HCl. The resultingmixture was stirred for 2 h and then extracted with ether (4×70 mL). Thecombined ether layers were washed with water (50 mL) and brine (50 μL),dried over Na₂SO₄, filtered, and evaporated. The residue was purified onsilica gel (eluant: 10% ether/hexanes) to yield two components. The morepolar component (200 mg) was the desired product. ¹H-NMR showed that itmight exist in a semi-ketal form. ¹H NMR (500 MHz, CDCl₃) δ 4.43-3.38(m, 5H), 3.24, 3.18 (ss, 3H) 2.52 (m, 1H), 1.82 (m, 1H). The less polarproduct (100 mg) was confirmed as alpha-alpha′ di-trifluoromethyltetrahydro-4H-pyran-4-one. ¹H NMR (500 MHz, CDCl₃) δ 4.59 (dd, 2H),3.24, 3.80 (t, J=11.3 Hz, 2H) 3.42 (m, 2H).

To a solution of 5-trifluoromethyl-2-pyridinal (51 g, 310 mmol) andsodium acetate (26.2 g, 319 mmol) in glacial acetic acid (200 mL) wasadded bromine (16.7 mL, 325 mmol) and the resulting mixture was heatedat 80° C. for 2.5 h. The reaction was allow to cool to room temperatureand then was evaporated under reduced pressure. The residue wasneutralized with saturated NaHCO₃ solution and extracted with ethylacetate (3×200 mL). The organics were combined, dried over MgSO₄,filtered, and evaporated in vacuo to yield 74.45 g (98%) of the crudeproduct. ¹H NMR (400 MHz, CDCl₃) δ 8.04 (d, J=2.6 Hz, 1H), 7.89 (m, 1H).

Under nitrogen, the substituted pyridine described in Step A,Intermediate 8 (48.8 g, 202 mmol) was added in small portions to asuspension of NaH (8.9 g, 220 mmol) in anhydrous tetrahydrofuran (500mL). After complete addition of the intermediate, the reaction mixturewas cooled to −78° C. and treated with tert-butyllithium (260 mL, 444mmol) added dropwise via syringe. After stirring for 5 min,N,N-dimethylformamide (50 mL, 707 mmol) was added slowly to maintain thetemperature below −50° C. The resulting mixture was then stirred for 10h allowing it to warm to room temperature. The mixture was quenched with2 N HCl and then diluted with ethyl acetate (1000 mL). The organic layerwas separated, washed with brine, dried over MgSO4, and evaporated invacuo. The desired product was precipitated out of ethyl acetate andhexanes and filtered to yield a light brown solid (28.55 g, 74%). ¹H NMR(500 MHz, CD₃OD) δ 10.13 (s, 1H), 8.21 (s, 2H).

A mixture of the intermediate from Step B, Intermediate 8 (18 g, 95mmol), sodium formate (7.1 g, 110 mmol), hydroxylamine hydrochloride(7.3 g, 110 mmol), and formic acid (150 mL) was stirred at roomtemperature for 2 h and then heated to reflux overnight. The reactionmixture was cooled and allowed to stand at room temperature for 7 days.The reaction was poured into water and extracted with ethyl acetate (3×). The combined organic layers were washed with water (2 ×), saturatedNaHCO₃ and brine, dried over Na₂SO₄, filtered, and concentrated in vacuoto yield the desired product as a brown powder (17.84 g, 90%). ¹H NMR(400 MHz, CD₃OD) δ 8.37 (d, J=2.7 Hz, 1H), 8.19 (q, J=0.7 Hz, 0.3 Hz,1H).

To a mixture of phosphorous oxychloride (13.4 mL, 144 mmol) andquinoline (8.7 mL, 73 mmol) was added the product from Step C,Intermediate 8, (24.6 g, 131 mmol) and the resulting mixture was heatedto reflux for 3 h. The reaction was cooled to 100° C. before water (70mL) was slowly added. The mixture was further cooled to room temperatureand neutralized carefully with saturated NaHCO₃ solution. The aqueouslayer was extracted with ethyl acetate (3 ×) and the organic layers werecombined, dried over MgSO₄, filtered, and evaporated in vacuo. The crudeproduct was purified by flash chromatography to afford (23.5 g, 87%) ofthe desired compound. ¹H NMR (500 MHz, CDCl₃) δ 8.88 (d, J=2.0 Hz, 1H),8.26 (d, J=2.5 Hz, 1H).

To a suspension of NaH (7.8 g, 200 mmol) in tetrahydrofuran (100 mL)under nitrogen was added dropwise a solution of tert-butyl methylmalonate (20 mL, 120 mmol) in anhydrous tetrahydrofuran (100 mL) viasyringe. The reaction mixture was stirred for 0.5 h before a solution ofthe intermediate prepared in Step D, Intermediate 8 (20.1 g, 97.6 mmol)in tetrahydrofuran (200 mL) was added slowly via syringe. The reactionwas stirred at room temperature overnight, then quenched with asaturated solution of NH₄Cl. The organic layer was separated and theaqueous layer was extracted with ethyl acetate (3 ×). The combinedorganic layers were washed with water (3 ×), dried over Na₂SO₄,filtered, and evaporated in vacuo. Flash chromatography afforded 31.76 g(95%) of the pure desired product. ¹H NMR (500 MHz, CDCl₃) δ 9.03 (d,J=1.5Hz, 1H), 8.25 (d, J=2.0Hz, 1H), 5.25 (s, 1H), 3.86(s, 3 H), 1.52(s, 9H).

A suspension of Raney Ni (1 g) and the product from Step E, Intermediate8 (18.2 g, 52.9 mmol) in ethanol (130 mL) was placed on a Parr apparatusand hydrogenated at 40 psi H₂ overnight. The suspension was filteredthrough celite and the filtrate was evaporated in vacuo to afford 16.35g (98%) of the crude product. ¹H NMR (500 MHz, CDCl₃) δ 8.83 (s, 1H),7.89 (s, 1H), 7.82 (s, 1H), 4.83 (d, J=16 Hz, 1H), 4.72 (s, 1H), 4.49(d, J=16 Hz, 1H), 1.45 (s, 9H).

To the mixture of the product from Step F, Intermediate 8 (16 g, 51mmol) in dichloromethane (60 mL) was added TFA (30 mL) and the resultingmixture was stirred at room temperature for 0.5 h. The solution wasevaporated under reduced pressure and the residue was dissolved indichloromethane. The mixture was neutralized by the slow addition of asolution of saturated sodium bicarbonate and the organic layer wasremoved. The aqueous layer was extracted with dichloromethane (4 ×) andthe combined organic layers were dried over Na₂SO₄, filtered, andevaporated in vacuo to afford 10.42 g (95%) of the desired product. ¹HNMR (400 MHz, CDCl₃) δ 8.81 (s, 1H), 7.78 (s, 1H), 7.30 (s, 1H), 4.63(s, 2H), 3.90 (s, 2H).

To a solution of the product from Step G, Intermediate 8 (18.0 g, 83.3mmol) in tetrahydrofuran (50 mL) was added 1.0 M borane intetrahydrofuran (417 mL, 420 mmol) and the resulting solution wasstirred at room temperature overnight. The solution was evaporated underreduced pressure and the residue was treated with 1% HCl/methanolsolution. The resulting mixture was heated at 50° C. overnight tobreakdown the borane complex. Treatment with acidic methanol wasrepeated twice to insure that the borane complex was removed. A solutionof this crude product (83.3 mmol, assuming 100% conversion) anddiisopropylethylamine (43 mL, 250 mmol) in dichloromethane was treatedwith di-tert-butyl dicarbonate (36.4 g, 167 mmol) and the resultingmixture was stirred at room temperature overnight. The solution waswashed with saturated sodium bicarbonate solution, water, and brine. Theaqueous layers were combined and back-washed with dichloromethane (2 ×).The combined organic layers were then dried over Na₂SO₄, filtered, andevaporated to dryness. The crude product was purified by flashchromatography and MPLC to afford (11.89 g, 47%) as a yellow solid. ¹HNMR (500 MHz, CDCl₃) δ 8.69 (s, 1H), 7.66 (s, 1H), 4.67 (s, 2H), 3.79(t, J=6.0 Hz, 2H), 3.08 (t, J=5.5 Hz, 2H), 1.51 (s, 9H).

The product described in Step H, Intermediate 8 (11.89 g) was treatedwith a solution of 4 N HCl in dioxane. The solution was stirred at roomtemperature for 2 h and then evaporated in vacuo to afford Intermediate8 (10.85 g, 99%) as a yellow powder. LC-MS for C₉H₁₀F₃N₂ calculated202.07, found [M+H]⁺203.0.

Procedure A:

A mixture of (1R,4S)-4-amino-cyclopen-2-ene carboxylic acid (130 g, 1.0mol), water (250 mL), sodium bicarbonate (170 g, 2.0 mol) andtetrahydrofuran (750 mL) was stirred for 30 min, then soliddi-tert-butyl dicarbonate (230 g, 1.05 mol) was added. The mixture wasstirred over the weekend, filtered to remove the insoluble material,evaporated to remove the tetrahydrofuran, and cooled to 0° C. To theresidue was added 2 N aqueous HCl until the pH reached 3 (˜500 mL). Theresulting precipitate was collected by filtration and washed with waterand dried under vacuum overnight. The desired acid was obtained as awhite solid (230 g, 100%). ¹H NMR (400 MHz, CD₃OD): δ 5.95 (m, 1H), 5.79(m, 1H), 4.80 (br s, 1H), 3.45 (m, 1H), 2.50 (m, 1H), 1.79 (m, 1H), 1.44(s, 9H).

The acid prepared in Step A (230 g, 1.0 mol) and 10% Pd/C (5.0 g) in 500mL of methanol was placed on a Parr apparatus and hydrogenated under 50psi of hydrogen for 1 h. The catalyst was removed by filtration and thefiltrate was evaporated. The residue was dissolved in dichloromethaneand dried over anhydrous sodium sulfate. After filtration, the filtratewas evaporated and dried under vacuum. The title compound was obtainedas a light yellow solid (230 g, 99%). LC-MS for C₁₁H₁₉NO₄calculated 229,found [M+H]⁺230.

To a mechanically stirred solution of the acid prepared in Step B,Procedure A, Intermediate 9 (230 g, 1.00 mol) in 500 mL ofN,N-dimethylformamide was added solid potassium carbonate (210 g, 1.5mol). The resulting mixture was stirred for 20 min and neat benzylbromide (120 mL, 1.0 mol) was added in one portion. An exothermicreaction was observed. After being stirred for 3 h at room temperature,the entire mixture was poured into an ice-water mixture (1000 mL). Thecrude product was extracted out with ether (2×800 mL). The combinedether layers were washed with water, dried over sodium sulfate, filteredand evaporated to offer a yellow solid. This solid was mixed with 4 NHCl in dioxane (400 mL), stirred overnight and condensed. The resultingsolid was collected by filtration, washed with ether and dried undervacuum. The title product was obtained as a hydrochloride salt (140 g,55%). ¹H NMR (400 MHz, CD₃ OD): δ 5.15 (s, 2H), 3.65 (m, 1H), 3.02 (q,J=8 Hz, 1H), 2.50 (m, 1H), 2.15 (m, 1H), 2.05 (m, 2H), 1.90 (m, 1H),1.75 (m, 1H).

The amino benzyl ester HCl salt prepared in Step C, Procedure A,Intermediate 9 (130 g, 0.50 mol) was suspended in 500 mL ofdichloromethane. Benzophenone imine (91 g, 0.50 mol) was added. Theresulting mixture was stirred overnight, and filtered to remove theinorganic salt. The filtrate was washed with water and brine, dried oversodium sulfate, and evaporated. The residue was dissolved in 200 mL oftoluene, and evaporated. This procedure was repeated once more. Thetitle compound (178 g) was obtained as a brown oil which was used in thenext step without further purification. ¹H NMR (400 MHz, CDCl₃): δ 1.80(m, 1H), 1.95 (m, 2H), 2.15 (m, 2H), 2.50 (m, 1H), 2.89 (m, 1H), 3.61(m, 1H), 5.20 (s, 2H), 7.18 (d, 2H), 7.38 (m, 8H), 7.47 (m, 3H), 7.64(d, 2H).

The starting Schiff base benzyl ester from Step D, Procedure A,Intermediate 9 (76.6 g, 200 mmol) in 300 mL of tetrahydrofuran wascooled to −78° C. under nitrogen. While stirring, a solution of lithiumdiisopropylamide (2.0 M, 110 mL, 220 mmol) in heptane was added over 20min. The mixture was stirred for 30 min at −78° C., then a solution of68 mL of isopropyl iodide (440 mmol) in 50 mL of tetrahydrofuran wasadded, and the mixture was allowed to stir for 30 min. The reactiontemperature was raised to 0° C. by removing the cooling bath. Afterbeing stirred for 2 h, the entire mixture was evaporated to remove thetetrahydrofuran. The residue was dissolved in ether (1000 mL), washedwith water and brine, dried over sodium sulfate, and evaporated. Thecrude product was dissolved in 500 mL of tetrahydrofuran, mixed with 400mL of aqueous 1N HCl, stirred for 1 h, and evaporated to removetetrahydrofuran at 50° C. The aqueous solution was extracted withhexanes (3 ×), made alkaline with saturated aqueous sodium carbonate(pH>9) and treated with a solution of di-tert-butyl dicarbonate (53 g)in 500 mL of dichloromethane. The resulting reaction mixture was stirredfor 30 min. The organic phase was separated and the aqueous phase wasextracted with dichloromethane (3 ×). The combined organic phases weredried over sodium sulfate and evaporated. The residue was purified byflash chromatography (silica gel, 10% ethyl acetate/hexanes) to yield amixture of the title compound as a mixture of cis and trans isomers(˜1:1, 24 g). Further purification by MPLC (8% ethyl acetate/hexanes)afforded the single desired cis isomer (fast-eluted, 7.3 g) and theundesired trans isomer (slow-eluted). ESI-MS calculated for C₂₁H₃₁NO₄:361; Found: [M+H]⁺362. ¹H NMR (500 MHz, CDCl₃): δ 7.36 (m, 5H), 5.14 (s,2H), 4.77 (m, 1H), 4.01 (d, J=5.0 Hz, 1H), 2.17 (m, 1H), 1.99-1.53 (m,5H), 1.42 (m, 9H), 0.85 (d, J=7.0 Hz, 6H).

The BOC-amine from Step E (7.3 g, 21 mmol) was treated with hydrogenchloride (4 N solution in dioxane). The reaction was allowed to stir for1.5 h at room temperature before being concentrated to remove thedioxane. The resultant solid was dissolved in dichloromethane (150 mL)and treated with tetrahydropyranone (2.4 g, 24 mmol) and triethylamine(2.8 mL, 20 mmol). The resulting solution was stirred at roomtemperature for 5 min before 4 Å powdered molecular sieves (˜5 g) andsodium triacetoxyborohydride (17 g, 80 mmol) where added. The mixturewas stirred for 2 h at room temperature. The reaction was filteredthrough celite and washed with a saturated aqueous sodium bicarbonatesolution then brine. The organic layer was dried over MgSO₄, filtered,and concentrated under reduced pressure. To give 6.7 g of a colorlessoil (97%). ESI-MS calculated for C₂₁H₃₁NO₃: 345; Found: 346 (M+H).

The amine from Step F (6.6 g, 19 mmol) was added to a solution ofdichloromethane (100 mL) and triethylamine (2.9 mL, 21 mmol).Trifluoroacetic anhydride (3.0 mL, 21 mmol) was added to the solutiondropwise at room temperature and the resulting solution was allowed tostir at room temperature for 2.5 h. The reaction was diluted withdichloromethane (100 mL) and washed with hydrochloric acid (1N aqueoussolution) followed by brine. The organic layer was dried over MgSO₄,filtered and concentrated under reduced pressure. The crude yellow oilwas purified by MPLC (silica gel, 0-30% ethyl acetate/hexanes) to give4.9 g of a colorless oil (58%). ¹H NMR (CDCl₃, 500 MHz): δ 7.37 (m, 5H),5.18 (m, 2H), 4.20-3.88 (m, 4H), 3.64 (m, 1H), 3.42 (t, J=12.0 Hz, 1H),3.26 (t, J=11.5 Hz, 1H), 3.18 (t, J=11.5 Hz, 1H), 2.81-2.65 (m, 2H),2.26 (m, 1H), 1.89-1.80 (m, 3H), 1.64-1.40 (m, 3H), 0.874 (m, 6H).

The product from Step G (3.5 g, 7.9 mmol) was dissolved in methanol (60mL) and treated with 20% palladium hydroxide on activated carbon (350mg). This mixture was placed under a hydrogen atmosphere (1 atm) andallowed to stir at room temperature for 1.2 h. The reaction was filteredthrough celite and concentrated under reduced pressure to give 2.63 g ofa white solid (95%).

Procedure B:

To a magnetically stirred solution of the acid from Step A, Procedure A,Intermediate 9 (159 g, 700 mmol) in 500 mL of N,N-dimethylformamide wasadded solid potassium carbonate (138 g, 1.00 mol). The resulting mixturewas stirred for 20 min before neat benzyl bromide (84 mL, 0.7 mol) wasadded in one portion. An exothermic reaction was observed. After stirredovernight at room temperature, the entire mixture was poured into anice-water mixture (1000 mL). The crude product was extracted out withethyl acetate (2×800 mL). The combined organic layers were washed withwater, dried over sodium sulfate, filtered and evaporated to offer abrown oil. This material was mixed with 4 N HCl in dioxane (350 mL) andstirred until gas evolution was observed. 500 mL of ether was added andthe precipitate was collected by filtration and washed with ether andhexanes. The desired product was obtained as a hydrochloride salt (164g, 93%). ¹H NMR (400 MHz, CD₃OD): δ 7.38 (m, 5H), 6.25 (m, 1H), 5.94 (m,1H), 5.20 (s, 2H), 4.32 (br s, 1H), 3.80 (br s, 1H), 2.67 (m, 1H), 2.14(m, 1H).

To a mixture of the amino ester HCl salt from Step A, Procedure B,Intermediate 9 (38 g, 150 mmol), tetrahydro-4-H-pyran-4-one (15 g, 150mmol), diisopropylethylamine (20.6 g, 160 mmol) and 4 Å powderedmolecular sieves (˜20 g) in 200 mL of dichloromethane was added sodiumtriacetoxyborohydride (42.4 g, 200 mmol) in multiple portions. Aftercomplete addition, the mixture was stirred at room temperatureovernight, quenched with saturated aqueous sodium carbonate, andfiltered through celite. The crude product was extracted intodichloromethane (3 ×), dried over sodium sulfate and evaporated. Theresidue was purified by flash chromatography (silica gel, 10%[aqueousNH₄ OH/methanol 1/9 ]/dichloromethane). The desired fractions werecombined and evaporated. The resulting residue was mixed withtetrahydrofuran and evaporated, redissolved in toluene and evaporated,and dried under vacuum to yield a light brown oil (38 g, 84%). ¹H NMR(400 MHz, CDCl₃): δ 7.38 (m, 5H), 5.98 (m, 1H), 5.85 (m, 1H), 3.98 (m,3H), 3.54 (m, 1H), 3.40 (m, 2H), 2.82 (m, 1H), 2.44 (m, 1H), 1.90 (m,1H), 1.79 (m, 2H), 1.70 (m, 1H), 1.44 (m, 2H).

To a round bottom flask containing solid potassiumbis(trimethylsilyl)amide (30 g, 150 mmol) under nitrogen was added 500mL of anhydrous tetrahydrofuran, and the resulting solution was cooledto −78° C. A solution of the amino ester from Step B, Procedure B,Intermediate 9 (38 g, 130 mmol) in 100 mL of tetrahydrofuran was addedover 20 min. The reaction mixture was warmed to −15° C. The mixture wasstirred at −15° C. for 1 h and then recooled to −78° C. A neat solutionof isopropyl iodide (65 mL, 380 mmol) was added. The flask was placedinto a −15° C. bath again. After a few min, a large amount of whiteprecipitate was formed. The reaction mixture was stirred for anadditional 1 h, poured into a mixture of ice and water, and extractedwith ethyl ether (3 ×). The ether layers were washed with water andbrine, dried over sodium sulfate and evaporated. The resulting residuewas dissolved in dichloromethane, dried over sodium sulfate again andevaporated. The residue was dried under vacuum, mixed withdichloromethane (200 mL) and cooled to 0° C. under nitrogen. To thesolution was added pyridine (33 mL, 400 mmol) and trifluoroaceticanhydride (27 mL, 190 mmol) dropwise. After 1 h, the reaction wasquenched with water. The organic phase was separated and washed with 2 Naqueous HCl, water and then brine. After being dried over sodium sulfateand evaporated, the residue was purified by flash chromatography (silicagel, 20% ethyl acetate/hexanes) to yield a light brown oil (41 g, 74%).¹H-NMR showed a 5:1 mixture of cis/trans isomers). ¹H NMR (400 MHz,CDCl₃): δ CH═CH: C is: 6.06 (m, 1H), 5.68 (m, 1H). Trans: 5.92 (m,0.2H), 5.79 (m, 0.2H). LC-MS for C₂₃H₂₇F₃NO₄ calculated 439, found[M+H]⁺440.

The unsaturated benzyl ester from Step C, Procedure B, Intermediate9(41g) and 10% Pd/C (2.0 g) in ethyl acetate (100 mL) was hydrogenated ona Parr apparatus under 50 psi of hydrogen overnight. The catalyst wasremoved by filtration through a pad of celite. The filtrate wasevaporated and dissolved in dichloromethane, evaporated and dried undervacuum overnight. The desired acid was obtained as a gummy white solid(32.5 g, 100%). LC-MS for C₁₆H₂₃F₃NO₄ calculated 351, found [M+H]⁺352.

To a solution of 2-hydroxy-3,5-dinitropyridine (5.0 g, 27 mmol) inN,N-dimethylformamide (15 mL) was added powdered potassium carbonate (54mmol) and the resulting mixture was stirred at 0° C. for 2 min. MethylIodide (27 mol) was then added slowly and the mixture was warmed up toroom temperature. After stirring for an additional 1 h, the reddishorange mixture was filtered and the filtrate was concentrated.Purification by column chromatography and eluting with hexanes/ethylacetate (0-50%) afforded 5.07 g (96%) of the desired product. ¹H NMR(500 MHz, DMSO): δ 9.59 (d, J=2.9 Hz, 1H), 9.00 (d, J=2.9 Hz, 1H), 3.67(m, 3H).

To the intermediate from Step A (4.0 g, 20 mmol) in 200 mL of 2 Mmethanol/ammonia was added 1-benzyl-4-piperdone (4.5 g, 24 mmol) and theresulting mixture was heated at 60° C. for 24 h. The solvent wasevaporated and the crude mixture was purified by flash columnchromatography. Eluting with hexanes/ethyl acetate (15-20%) gave 4.0 g(72%) of the title product. ¹H NMR (500 MHz, CD₃OD): δ 9.14 (d, J=2.5Hz, 1H), 8.30 (d, J=2.3 Hz, 1H), 7.40-7.28 (m, 5H), 3.76 (s, 4H), 3.10(t, J=6.0 Hz, 2H), 2.91 (t, J=6.0 Hz, 2H).

A mixture of the intermediate from Step B (4 g) and Pd/C (250 mg, 5%) inmethanol (125 mL) was hydrogenated at room temperature for 3.5 h. Themixture was filtered through celite and concentrated. Purification bycolumn chromatography and eluting with hexanes/ethyl acetate (1:1) andmethanol (5%) afforded 2.52 g (71%) of the title product. ¹H NMR (500MHz, CD₃OD): δ 7.79 (d, J=2.5 Hz, 1H), 7.39-7.28 (m, 5H), 6.76 (d, J=2.5Hz, 1H), 3.69 (s, 2H), 3.53 (s, 2H), 2.84-2.81 (m, 4H).

To a mixture of the intermediate from Step C (2.4 g, 10 mmol) and 10 mLof 20% sulfuric acid at 0° C. was added a solution of sodium nitrite(0.76 g, 11 mmol) in water (5 mL). After stirring at 0° C. for 15 min, asmall crystal of urea was added and the resulting mixture was addedslowly to 20% sulfuric acid (85 mL) at 90° C. Heating was continued foran additional 30 min, the mixture was cooled, and the pH was adjusted to7 with potassium carbonate (solid). The mixture was extracted withdichloromethane (2×100 mL) and the organic layer was washed with brine,dried (MgSO₄) and concentrated in vacuo. Purification by columnchromatography and eluting with hexanes/ethyl acetate (1:1)+3% methanol,afforded 1.44 g (60%) of the title product. ¹H NMR (500 MHz, CD₃OD): δ7.88 (d, J=2.5 Hz, 1H), 7.39-7.28 (m, 5H), 6.89 (d, J=2.5 Hz, 1H), 3.70(s, 2H), 3.58 (s, 2H), 2.89-2.81 (m, 4H).

A mixture of the intermediate from Step D (1.45 g), ethanol (25 mL), 2 NHCl (5.0 mL) and Pd/C (100 mg, 10%) was hydrogenated at room temperaturefor 24 h and the resulting mixture was filtered through celite. Thecatalyst was washed thoroughly with hot ethanol and the filtrate wasconcentrated in vacuo to yield 1.2 g of the desired product as the HClsalt. ¹H NMR (500 MHz, CD₃OD): δ 8.29 (d, J=2.6 Hz, 1H), 7.87 (d, J=2.3Hz, 1H), 4.57 (s, 2H), 3.68 (t, J=6.5 Hz, 2H), 3.37 (t, J=6.2 Hz, 2H).

To a solution of the amine intermediate from Step E (1.20 g, 5.3 mmol)in 40 mL of water/dichloromethane (1:1) was added di-tert-butyldicarbonate (1.40 g) followed by sodium bicarbonate (2.25 g). Themixture was stirred vigorously at room temperature for 4 h. The layerswere separated and the aqueous layer was washed with dichloromethane(×2). The combined dichloromethane layers were dried (MgSO₄),concentrated and chromatographed. Eluting with hexanes/ethyl acetate(1:1)+5% methanol gave 0.91 g (68%) of the title product ¹H NMR (500MHz, CD₃OD): δ 7.91 (d, J=2.5 Hz, 1H), 7.01 (d, J=2.9 Hz, 1H), 4.53 (s,2H), 3.72 (t, J=5.5 Hz, 2H), 2.83 (t, J=6.0 Hz, 2H), 1.48 (s, 9H).

Procedure A:

A mixture of (1S)-(+)-2-azabicyclo[2.2.1]hept-5-en-3-one (10.3 g, 94.4mmol) in ethyl acetate (200 mL) and 10% Pd/C (0.5 g), was hydrogenatedat room temperature. After 24 h the reaction mixture was filtered andevaporated leaving behind 10.4 g (100%) of the product that was taken in250 mL methanol and HCl (12 M, 6 mL). The resultant mixture was stirredat room temperature, until the reaction was complete (72 h). Evaporationof methanol followed by drying under high vacuum, yielded title compoundas an off white solid (16.0 g, 96%). ¹H NMR (500 MHz, D₂O): δ 3.70 (s,3H), 3.01 (m, 1H), 2.38 (m, 1H), 2.16-1.73 (m, 6H).

To a suspension of the intermediate from Step A (10.2 g, 56.8 mmol) indry dichloromethane (200 mL) was added benzophenone imine (10.2 g, 56.8mmol) at room temperature and the resultant mixture was stirred for 24h. The reaction mixture was filtered and the filtrate was evaporated, toleave behind a yellow oil that was triturated with ether (100 mL),filtered and evaporated. This operation was repeated twice to ensurethat the product was free of ammonium chloride impurities. The resultantoil was thoroughly dried under vacuum to yield the title compound (18.03g, >100%) and required no further purification. 1H NMR (500 MHz, CDCl₃):δ 7.5-7.18 (m, 10H), 3.75 (m, 1H), 3.7 (s, 3H), 2.78 (m, 1H), 2.26-1.71(m, 6H).

To a solution of lithium diisopropylamide (prepared fromdiisopropylamine (7.7 g, 76 mmol) and n-butyllithium (30.4 mL, 2.5 M inhexanes, 76 mmol) in tetrahydrofuran (120 mL) at −78° C. was added theester from step B (18.0 g, 58.6 mmol). The resultant burgundy coloredsolution was stirred for 20 min after which it was quenched with2-iodopropane (14.9 gm, 88 mmol). The reaction mixture was graduallywarmed over 3 h to 0° C. and this temperature was maintained for anadditional 3 h. Reaction was quenched with water and extracted withethyl acetate. The organic layer was washed with water, brine, dried(anhydrous magnesium sulfate) and concentrated to yield an oil. To thesolution of the crude Schiff base (20.0 g) in tetrahydrofuran (100 mL)was added HCl (5.0 mL, 12 M). The resulting reaction mixture was allowedto stir at room temperature for 3 h. After the removal of all volatiles,the hydrochloride salt was taken up into dichloromethane (250 mL),saturated solution of sodium bicarbonate (250 mL) and di-tert-butyldicarbonate (26.0 g, 1.4 Eq.) were added. The resultant mixture wasvigorously stirred overnight at room temperature. The organic layer wasseparated and washed with water, brine, dried (anhydrous magnesiumsulfate) and concentrated to yield an oil. Purification by flash columnchromatography (eluent: hexanes/ethyl acetate 19:1) gave the desiredproduct (4.91 g, 30%). ¹H NMR (500 MHz, CDCl₃): 4.79 (br, 1H), 4.01 (m,1H), 3.71 (s, 3H), 2.18-1.60 (m, 6H), 1.44 (s, 9 H), 0.87 (d, J=6.9 Hz,3 H), 0.86 (d, J=6.9 Hz, 3H).

To a solution of the ester from Step C (4.91 g, 17.2 mmol) in methanol(100 mL) was added a solution of LiOH (3.6 g, 85 mmol) in water (20 mL)and tetrahydrofuran (10 mL). The resultant mixture was heated at 80° C.until the reaction was complete (18 h). The methanol was removed invacuo and the crude product was taken up with water/ethyl acetate (200mL, 1:4) and cooled to 0° C. The acidity of the mixture was adjusted topH 6. The ethyl acetate layer was separated, washed with water, brine,dried (anhydrous magnesium sulfate) and concentrated to yield an oil.Purification by flash column chromatography (eluent: hexanes/ethylacetate 1:1+2% AcOH) gave Intermediate 11 (3.9 g, 84%). 1H NMR (500 MHz,CDCl₃): 11.36 (br, 1H), 6.49 (br, 1H), 4.83 (m, 1H), 3.71 (s, 3H),2.30-1.55 (m, 6H), 1.46 (s, 9H), 0.94 (d, J=6.9 Hz, 3H), 0.933 (d, J=6.9Hz, 3H).

Procedure B:

Commercially available (1R,4S)₄-aminocyclopent-2-ene-1-carboxylic acidwas converted to its methyl ester hydrochloride salt via classicalprocedures.

To a suspension of amine from Step A (6.31 g, 35.5 mmol) in acetone (40mL) and water (20 mL) was added solid NaHCO₃ (6.6 g, 78 mmol) inportions. After 5 min, a solution of di-tert-butyl dicarbonate (8.5 g,39 mmol) in acetone (60 mL) was added and the reaction mixture wasstirred at room temperature. After 3 h, acetone was removed in vacuo andthe residue was partitioned between ether (500 mL) and saturated aqueousNaHCO₃ solution (120 mL). The ether layer was further washed withaqueous NaHCO₃ solution (1×100 mL), brine (1×100 mL), dried overanhydrous Na₂SO₄, concentrated and purified by flash chromatography (15%ethyl acetate/hexanes) to afford the product (7.25 g, 85%).

To a solution of lithium bis(trimethylsilyl)amide (10.4 g, 62.1 mmol) intetrahydrofuran (100 mL) was added a solution of the intermediate fromStep B (6.71 g, 27.8 mmol) in tetrahydrofuran (10 mL) over 10 min at−78° C. The resulted solution was stirred at −78° C. for 30 min beforeisopropyl iodide (3.3 mL, 33 mmol) was added in one portion. Thereaction was allowed to warm up to −25° C. and this temperature wasmaintained overnight. The reaction was then quenched with an aqueoussaturated NH₄Cl solution (250 mL). The organic layer was separated andthe aqueous layer was further extracted with diethyl ether (3×100 mL).The combined organic layers were then washed with brine (1×100 mL),dried over anhydrous Na₂SO₄, filtered, concentrated and purified byflash chromatography (5-10% ethyl acetate/hexanes) to give the product(5.66 g, 72%) as a clear oil (cis/trans=4.3/1). ¹H NMR (500 MHz, CDCl₃)cis-isomer: δ 5.79 (s, 2H), 4.75 (m, 1H), 3.72 (s, 3H), 2.28-2.20 (m,2H), 2.0 (dd, J=15, 4 Hz, 1H), 1.45 (s, 9H), 0.85 (d, J=6.6 Hz, 3H),0.81 (d, J=7 Hz, 3H).

To a solution of the product from step C (1.6 g, 5.7 mmol) intetrahydrofuran (50 mL), methanol (50 mL) and water (10 mL) was addedLiOH monohydrate (400 mg) and the reaction was heated to refluxovernight until the TLC indicated that the reaction was complete. Theorganic solvents were removed in vacuo and the aqueous layer was washedwith ether (1×) and then acidified slowly with concentrated HCl untilthe pH reached 4. The resulting suspension was extracted with CH₂Cl₂ (3×). The combined organic layers were dried over anhydrous MgSO₄,filtered and concentrated to give the product as a mixture of twocis/trans isomers (1.5 g) as a foaming yellow solid. This solid wasdissolved in ethyl acetate (2 mL) with heating and diluted with hexanes(50 mL) to give a clear solution. This solution was allowed to cool toroom temperate slowly over 1 h and then maintained at −25° C. in afreezer overnight. The trans-isomer was crystallized out along with someof the desired cis-isomer (500 mg total). The mother solution wascollected and concentrated to give the title compound (1 g, 66%,cis-isomer only). ¹H NMR (500 MHz, CDCl₃) cis-isomer: δ 5.80 (m, 2H),4.80 (m, 1H), 2.40-2.20 (m, 2H), 2.15-2.0 (m, 1H), 1.5 (m, 9H), 1.0-0.8(m, 3H).

To a solution of the product from Step D (1 g) in ethanol (30 mL) wasadded 10% Pd/C (100 mg) and the resulting mixture was agitated on a Parrapparatus at 50 lb pressure of H2 overnight. The mixture was filteredthrough celite and concentrated in vacuo to afford the title compound (1g, 99%). 1H NMR (500 MHz, CDCl3): 11.36 (br, 1H), 6.49 (br, 1H), 4.83(m, 1H), 3.71 (s, 3H), 2.30-1.55 (m, 6H), 1.46 (s, 9H), 0.94 (d, J=6.9Hz, 3H), 0.933 (d, J=6.9 Hz, 3H).

A flame dried 1000 mL round bottom flask was charged with 400 mL of drytetrahydrofuran, and then, set under nitrogen and cooled to −78° C.using an acetone/dry ice bath. Diisopropylamine (27.4 mL, 195 mmol) wasadded to the cooled solvent via a syringe. The resulting solution wasslowly treated with 2.5 M n-butyllithium in hexanes (55 mL, 140 mmol).After 5 min stirring, the product described in Step B, Intermediate 11(40 g, 130 mmol) in 100 mL of tetrahydrofuran was added dropwise viasyringe and the resulting mixture was stirred at −78° C. for 2 h.2-iodo-1,1,1 -trifluoroethane (47 mL, 480 mmol) was then added dropwisevia syringe and the resulting mixture was stirred overnight allowing itto warm slowly to room temperature. The reaction was quenched with asaturated solution of ammonium chloride (400 mL) and the organics wereseparated. The aqueous layer was extracted with ethyl acetate (3×150 mL)and all the organics were combined, dried over anhydrous sodium sulfate,filtered, and evaporated under reduced pressure. The crude product wasused in the next step without further purification. LC-MS forC₂₂H₂₂F₃NO₂ calculated 389.26, found [M+H⁺] 390.4

To a solution of the product from Step A, Intermediate 12 (130 mmol,assuming 100% conversion) in 200 mL of tetrahydrofuran was added 200 mLof 2 N hydrochloric acid and the resulting mixture was stirred overnightat room temperature. The solution was concentrate in vacuo to remove thetetrahydrofuran and the aqueous layer was then diluted withdichloromethane (300 mL). The pH of the aqueous layer was adjusted to apH of 10 by the slow addition of 5 N sodium hydroxide with vigorousstirring. The organic layer was removed using a separatory funnel andthe aqueous layer was extracted with dichloromethane (2×150 mL). Theorganic layers were combined, dried over anhydrous sodium sulfate, andfiltered. To the filtrate was added diisopropylethylamine (22.7 mL, 130mmol) and di-tert-butyl dicarbonate (32.7 g, 150 mmol) and the resultingsolution was stirred at room temperature overnight. The mixture waswashed with 1N hydrochloric acid, followed by a saturated solution ofsodium bicarbonate, and brine. The organic layer was dried overanhydrous sodium sulfate, filtered, and evaporated under reducedpressure. Purification by MPLC (5 g per run) afforded 5.87 g (14%) ofthe desired cis (R, S) isomer and 12.31 g (29%) of the undesired trans(S, S) isomer. Also, 5.22 g (12%)was recovered as a 1:1 mixture of the 2diastereomers. ¹H NMR (500 MHz, CDCl₃) δ (1^(st) desired isomer) 5.05and 4.40 (singlets, 1H), 3.76 (s, 3H), 2.73 (ddd, J=11.0, 12.8, 14.8 Hz,1H), 2.38 (ddd, J=10.7, 12.8, 15.0 Hz, 1H) 2.32-2.26 (m, 1H), 2.21 (brdd, J=3.6, 14.5 Hz, 1H), 2.18-2.11 (m, 1H), 2.02 (dd, J=8.8, 14.4 Hz,1H), 1.61 (dd, J=7.8, 13.2 Hz, 1H) 1.52 (br s, 10H). ¹H NMR (500 MHz,CDCl₃) δ (2^(nd) undesired isomer) 4.52 and 4.06 (singlets, 1H), 3.72(s, 3H), 2.72 (dd, J=7.1, 13.5 Hz, 1H), 2.66 (ddd, J=10.6, 12.8, 15.0Hz, 1H), 2.53 (ddd, J=11.0, 12.8, 14.9 Hz, 1H) 2.26 (app dd, J=7.1, 13.5Hz, 1H), 2.18-2.07 (m, 1H), 1.78 (dd, J=8.6, 13.5 Hz, 1H), 1.57-1.48 (m,2H) 1.46 (s, 9H).

To a mixture of the desired cis (R,S) product described in Step B,Intermediate 12 (4.0 g, 12 mmol) in a 1:1:1 solution oftetrahydrofuran/methanol/water (84 mL) was added solid LiOH (2.60 g,62.0 mmol) and the resulting solution was heated to 60° C. and stirredfor 18 h. The mixture was left standing to cool to room temperature andthen concentrated to remove the organic solvent. The aqueous layer wasacidified by the slow addition of 6 N hydrochloric acid to pH 4-5. Theacidic aqueous layer was extracted with dichloromethane (3×100 mL) andthe organics were combined, dried over anhydrous sodium sulfate,filtered, and evaporated under reduced pressure to afford Intermediate12 (3.86 g, 99%) as a yellow oil. After two days standing at 5° C. inthe refrigerator, the material crystallized.

This intermediate was prepared in an analogous fashion to Intermediate12, except 2 -iodo-1,1,1-trifluoroethane was replaced with3-iodo-1,1,1-trifluoropropane. Purification by MPLC (gradient eluant0-40% ethyl acetate/hexanes) afforded 612 mg (11%) of the desired cis(R, S) isomer (Intermediate 13) and 905 g (17%) of the undesired trans(S, S) isomer.

This intermediate was prepared in an analogous fashion to Intermediate12, except 2 -iodo-1,1,1-trifluoroethane was replaced with cyclobutylbromide. Purification by MPLC (gradient eluant 0-30% ethylacetate/hexanes) afforded 103 mg (5%) of the desired cis (R, S) isomer(Intermediate 14). The more polar trans isomer was not collected. ¹H NMR(500 MHz, CDCl₃) δ 4.85 and 4.10 (singlets, 1H), 2.28-2.21 (m, 1H), 2.13(dd, J=5.0, 14.0 Hz, 1H) 2.10-2.04 (m, 1H), 1.99 (dd, J=8.0, 13.7 Hz,1H), 1.68-1.56 (m, 2 H), 1.53 (dd, J=7.2, 13.6 Hz, 1H), 1.46 (br s, 10H), 0.64-0.56 (m, 1H), 0.46-0.37 (m, 2H), 0.08-0.01 (m, 2H).

This intermediate was prepared in an analogous fashion to Intermediate12, except 2-iodo-1,1,1-trifluoroethane was replaced withiodocyclopropane. Purification by MPLC (gradient eluant of 0-25% ethylacetate/hexanes) afforded 506 mg (20%) of the desired cis (R, S) isomer(Intermediate 15) and 803 g (32%) of the undesired trans (S, S) isomer.¹H NMR (500 MHz, CDCL3) δ ¹H NMR (500 MHz, CDCL3) δ (1^(st) desiredisomer) 4.80 and 4.02 (singlets, 1H), 2.27 (ddd, J=8.0, 9.7, 17.8 Hz,1H), 2.19 (ddd, J=4.4, 7.4, 12.4 Hz, 1H) 2.07-1.96 (m, 3H), 1.95 (br dd,J=8.2, 14.0 Hz, 1H) 1.68-1.50 (m, 8H), 1.45 (br s, 10H), 1.25-1.17 (m,1H). ¹H NMR (500 MHz, CDCl₃) δ (2^(nd) undesired isomer) 4.56 and 3.90(singlets, 1H), 2.58 (dd, J=7.1, 13.0 Hz, 1H), 2.22 (ddd, J=8.0, 9.6,17.7 Hz, 1H), 2.11 (ddd, J=7.5, 7.6, 13.3 Hz, 1H) 2.04-1.93 (m, 1H),1.68-1.45 (m, 7H), 1.44 (br s, 10H), 1.38-1.15 (m, 4H).

A solution of diisopropylamine (2.70 mL, 19.3 mmol) in tetrahydrofuran(20 mL) was cooled to −78° C. and a solution of n-butyllithium inhexanes (7.70 mL, 2.5 M, 19.3 mmol) was added via syringe, followed by asolution of the Schiff base, from Step C, Intermediate 9 (5.685 g, 14.82mmol) in tetrahydrofuran (10 mL). The enolate was allowed to form for 3h at −78° C., after which time the neat acetaldehyde (1.00 mL, 29.7mmol) was added. The reaction was quenched with the addition of aqueouscitric acid (200 mL, 10%) and the crude product was extracted intodiethyl ether. Drying (anhydrous magnesium sulfate) and evaporation ofthe solvent gave the crude desired product (6.16 g). This was furtherpurified by flash chromatography (deactivated silica gel, ethylacetate/hexanes 3:7) to yield the desired cis-isomer (2.32 g, 54%). ThisSchiff base was found to be unstable, and was used in the next stepwithout delay. LC-MS for C₂₈H₂₉NO₃ calculated: 427.21, found[M+H]⁺428.20.

The Schiff base from Step A (2.323 g, 5.433 mmol) was dissolved intetrahydrofuran (20 mL) and 2 N HCl was added. The reaction mixture wasstirred at room temperature for 2 h, after which time the volatiles wereremoved in vacuo. The resulting mixture of the desired aminehydrochloride and benzophenone was used in the next step without furtherpurification.

The crude product from the previous step (max 5.433 mmol) was dissolvedin dichloromethane (50 mL), di-tert-butyl dicarbonate (2.371 g, 10.87mmol) was added followed by 50 mL of a saturated solution of sodiumbicarbonate. The reaction mixture was vigorously stirred at roomtemperature for 1 h. The layers were separated and the aqueous phase waswashed with dichloromethane. The combined organic extracts were dried(anhydrous magnesium sulfate) and the solvent was evaporated in vacuo.Final purification by gradient flash chromatography (ethylacetate/hexanes 0-40%) gave the desired BOC-protected amine (619 mg,32%, two steps) as a mixture (3:2) of two diasteromers. LC-MS forC₂₀H₂₉NO₅ calculated: 363.20, found 264.20 ([M+H]⁺-loss of the BOCgroup).

This acid was prepared following the procedure described in Intermediate12, Step C, and was used in the next step without further purification.

A solution of the acid from the previous step (809 mg, 2.96 mmol),Intermediate 8 (1.63 g, 5.92 mmol), 1-hydroxy-7-azobenzotriazole (402mg, 2.96 mmol), and diisopropylethylamine (1.0 mL, 5.9 mmol) indichloromethane (25 mL) was treated with1-(-3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.70 g,8.88 mmol) and the resulting mixture was stirred at room temperatureovernight. The reaction was quenched with water, and the product wasextracted into dichloromethane. The combined organic extracts were dried(anhydrous magnesium sulfate) and the solvent was removed in vacuo. Theresidue (679 mg) was separated by MPLC (eluant gradient 40-100% ethylacetate/hexanes) to yield a single isomers (the hydroxyethyl side-chain)of unknown absolute stereochemistry. ¹H NMR (CDCl₃, 500 MHz) indicated amixture of isomeric alcohols in a ratio of about 2 to 3. LC-MS forC₂₂H₃₀F₃N₃O₄ calculated: 457.22, found 358.20 ([M+H]⁺-loss of the BOCgroup).

The solution of the of the higher eluting diastereoisomer from theprevious step (282 mg, 0.618 mmol) in dichloromethane (6 mL) was treatedwith TFA (4 mL) and the resulting mixture was stirred at roomtemperature for 2 h. The volatiles were removed in vacuo to yield 218 mg(99%) of the crude product. LC-MS for C₁₇H₂₂F₃N₃O₂ calculated: 357.17,found [M+H]⁺358.10.

A flame dried round bottom flask was charged with NaH (15 mg, 60%suspension, 0.4 mmol) and set under static nitrogen.N,N-dimethylformamide (2.0 mL) was added via syringe, and the mixturewas cooled to 0° C. While stirring, a solution of the benzyl ester fromStep A, Intermediate 16 (higher eluting (1,3-cis-) diastereoisomericpair, 142 mg, 0.332 mmol) and methyl iodide (142 μL, 1.00 mmol) wereadded via syringe. The cooling bath was removed and the mixture wasstirred at room temperature for 3 h. The reaction was quenched bypouring onto water and the crude product was extracted with a mixture ofhexanes and ether (1:1). The combined organic extracts were backwashedwith water, dried (anhydrous sodium sulfate) and the solvent wasevaporated in vacuo to leave 106.3 mg (73%) of crude product. The tworespective diastereoisomers were separated by gradient flashchromatography (eluent: 0-40% of ethyl acetate/hexanes). LC-MS forC₂₉H₃₁NO₃ calculated 441.23, found [M+H]⁺442.30.

This amine was synthesized starting from the product of Step A in aseries of reactions analogous to those described in Intermediate 16,Steps B-F.

This intermediate was prepared in an analogous fashion to Intermediate16, except acetaldehyde was replaced with propionaldehyde. Purificationby MPLC (gradient eluant 40-100% ethyl acetate/hexanes) afforded singleisomers (the hydroxypropyl side-chain) of unknown absolutestereochemistry (total yield of all 312 mg, 41%). Isomer 1: ¹H NMR (500MHz, CDCl₃) δ 5.0 (br s, 1H), 4.08 (br. S, 1H), 3.60 (ddd, J=2.0, 7.9,9.8 Hz, 1H), 2.50-2.42 (m, 2H), 2.10-1.88 (m, 4H), 1.64-1.52 (m, 2H),1.45 (s overlapped, 9H),1.65 (s, 1H) 1.48-1.36 (m, 1H), 1.29-1.22 (m,1H), 0.98 (t, J=7.3 Hz, 3H). Isomer 2: ¹H NMR (500 MHz, CDCl₃) δ 4.76(br s, 1H), 4.08 (br s, 1H), 3.63-3.55 (m, 1H), 2.26 (dd, J=7.8, 14.0Hz, 1H), 2.22-2.15 (m, 1H), 2.06-1.94 (m, 2H), 1.91 (dd, J=5.4, 14.1 Hz,1H), 1.76-1.68 (m, 1H), 1.60 (s, overlapped, 1H), 1.60-1.50 (m, 2H),1.45 (s, overlapped, 9H), 1.48-1.38 (m, 1H), 1.30-1.20 (m, 1H), 0.98 (t,J=7.2 Hz, 3H). Isomer 3: ¹H NMR (500 MHz, CDCl₃) δ 4.82 (br s, 1H), 4.09(br s, 1H), 3.43 (d, J=9.8 Hz, 1H), 2.19 (s, 1H), 2.11 (ddd, J=4.8, 7.2,12.7 Hz, 1H), 2.06-1.90 (m, 6H), 1.45 (s, overlapped, 9H), 1.54-1.40 (m,1H), 1.28-1.18 (m, 1H), 0.99 (t, J=7.1 Hz, 3H). Isomer 4: ¹H NMR (500MHz, CDCl₃) δ 4.83 (br s, 1H), 4.04 (br s, 1H), 3.59 (app br t, J=8.1Hz, 1H), 2.55 (br dd, J=7.1, 13.7 Hz, 1H), 2.39 (br d, J=7.1 Hz, 1H),2.18 (s, 1H), 2.14-2.06 (m, 1H), 2.02-1.91 (m, 2H), 1.90-1.82(m, 1H),1.72-1.65 (m, 1H), 1.59-1.50 (m, 1H), 1.44 (s, overlapped, 9H),1.47-1.37 (m, 1H), 1.26-1.17 (m, 1H), 0.96 (t, J=7.3 Hz, 3H).

Intermediate 8 (4.6 g, 16 mmol) and Intermediate 11 (4.0 g, 14 mmol)were first dried by azeotropic distillation with toluene (3×50 mL) andplaced under high vacuum for 30 min. Under nitrogen,4-dimethylaminopyridine (1.08 g, 8.60 mmol), anhydrous dichloromethane(40 mL), and diisopropylethylamine (7.0 mL, 40 mmol) were addedsequentially. After Intermediate 8 was in solution,bromo-tris-pyrrolidino-phosphonium hexafluorophosphate (6.80 g, 14.3mmol) was added, immediately followed by additionaldiisopropylethylamine (7.0 mL, 40 mmol). The reaction mixture wasstirred at room temperature overnight and then quenched with saturatedNaHCO₃. The aqueous layer was back washed with dichloromethane (3×50 mL)and the organic layers were combined, dried over Na₂SO₄, filtered, andevaporated in vacuo. The crude product was purified by flashchromatography (stepwise gradient 0-60%, ethyl acetate/hexanes) toafford the product (4.80 g, 74%) as a yellow foam. ¹H NMR (500 MHz,CDCL₃) δ 8.72 (s, 1H), 7.70 (s, 1H), 4.88 (br d, J=17.0 Hz, 1H), 4.78(d, J=17.6 Hz, 1H), 4.04-3.84 (m, 2H), 3.52 (br s, 1H), 3.12 (br t,J=5.6 Hz, 1H), 2.32-2.06 (m, 3H), 1.98-1.70 (m, 4H), 1.64-1.54 (m, 1H),1.44 (s, 9H), 0.92-0.82 (m, 6H). LC-MS for C₂₃H₃₂F₃N₃O₃ calculated455.24, found [M+H]⁺456.2.

The from Step B, Intermediate 19 (1.2 g, 2.6 mmol) was dissolved with 4N HCl in dioxane (50 mL) and the resulting solution was stirred at roomtemperature for 1 h. The reaction was evaporated under vacuum to affordthe product (904 mg, 97%) as a white powder. LC-MS calculated forC₁₈H₂₄F₃N₃O is 355.20, found [M+H]⁺356.2.

To a solution of the product described in Step A, Intermediate 19 (2.0g, 4.4 mmol) in dichloromethane (80 mL) was added 3-chloroperoxybenzoicacid (2.11 g, 8.83 mmol) and the resulting solution was stirredovernight at room temperature. The mixture was cooled to 0° C. and whilestirring vigorously, solid calcium hydroxide was added in portions(about 6 g). The suspension was stirred for an additional 30 min, thenfiltered through celite to remove all solids. The filtrate wasevaporated in vacuo and the residue was purified by MPLC (gradienteluant 40-100% ethyl acetate/hexanes) to afford 1.32 g (64%) of thedesired compound. ¹H NMR (500 MHz, CDCL₃) δ 8.46 (s, 1H), 7.28 (s, 1H),4.88 (br d, J=17.2 Hz, 1H), 4.78 (d, J=17.7 Hz, 1H), 4.05-3.84 (m, 2H),3.12 (br s, 1H), 2.34-2.06 (m, 3H), 1.88-1.70 (m, 4H), 1.62-1.54 (m,1H), 1.43 (s, 9H), 0.90-0.85 (m, 6H). LC-MS for C₂₃H₃₂F₃N₃O₅ calculated471.20, found [M+H]⁺472.2.

The product from Step B, Intermediate 20 (1.32 g, 2.82 mmol) wasdissolved in 4 N HCl in dioxane (50 mL) and the resulting solution wasstirred at room temperature for 1 h. The reaction was evaporated undervacuum to afford the product (1.10 g, 98%) as a white powder. LC-MS forC₁₈H₂₄F₃N₃O₂ calculated 371.20, found [M+H]⁺372.2.

Thionyl chloride (20.1 mL, 275 mmol) was slowly introduced to 175 mL ofmethanol and the resulting solution was allowed to stir for 10 min. Tothis solution, (1R,4S)-4-amino-cyclopent-2-ene (10 g, 79 mmol) was addedand the mixture was heated to reflux for 15 h. After allowing to cool toroom temperature, the solution was evaporated in vacuo to afford thecrude product (13.95 g, 99%) which was used in the next step withoutfurther purification.

To a suspension of the intermediate from Step A (13.9 g, 78.8 mmol) indry dichloromethane (100 mL) was added benzophenone imine (13.5 g, 78.5mmol) at room temperature and the resultant mixture was stirred for 24h. The reaction mixture was filtered and the filtrate was evaporated, toleave behind a yellow oil that was triturated with ether (100 mL),filtered and evaporated. This operation was repeated twice to ensurethat the product was free of ammonium chloride impurities. The resultantoil was thoroughly dried under high vacuum to yield the title compound(18.03 g, >100%) and required no further purification. ¹H NMR (500 MHz,CDCl₃): δ 7.64 (d, J=7.1 Hz, 2H), 7.52-7.44 (m, 3H), 7.38 (t, J=7.1 Hz,1H), 7.33 (t, J=7.1 Hz, 2H), 7.20 (d, J=7.1 Hz, 2H), 5.97 (ddd, J=2.1,4.1, 5.7 Hz, 1H), 5.78 (ddd, J=2.3, 4.8, 5.5 Hz, 1H), 4.52 (br ddd,J=2.1, 5.3, 7.3 Hz, 1H), 3.74 (s, 3H), 3.52 (ddd, J=2.2, 5.95, 8.4 Hz,1H), 2.40-2.33 (m, 1H), 2.29-2.22 (m, 1H).

A flame dried 500 mL round bottom flask was charged with 100 mL of drytetrahydrofuran, and then, set under nitrogen and cooled to −78° C.using an acetone/dry ice bath. Diisopropylamine (2.74 mL, 19.5 mmol) wasadded to the cooled solvent via syringe. 2.5 M n-butyllithium in hexanes(7.80 mL, 19.50 mmol) was then added slowly to the solution. After 5 minstirring, the product described in Step B. Intermediate 21 (5.0 g, 16mmol) in 30 mL of tetrahydrofuran was added dropwise via syringe and theresulting mixture was stirred at −78° C. for 2 h. 2-iodopropane (2.26mL, 22.8 mmol) was then added dropwise via syringe and the resultingmixture was stirred overnight allowing it to warm slowly to roomtemperature. The reaction was quenched with a saturated solution ofammonium chloride (100 mL) and the organic layer was separated. Theaqueous layer was extracted with ethyl acetate (3×100 mL) and all theorganics were combined, dried over anhydrous sodium sulfate, filtered,and evaporated under reduced pressure. The crude product was used in thenext step without further purification. LC-MS for C₂₃H₂₅NO₂ calculated347.19, found [M+H]⁺348.2.

To a solution of the product from Step C, Intermediate 21 (16.25 mmol,assuming 100% conversion) in 100 mL tetrahydrofuran was added 100 mL of2 N hydrochloric acid and the resulting mixture was stirred overnight atroom temperature. The solution was concentrate in vacuo to remove thetetrahydrofuran and the aqueous layer was then diluted withdichloromethane (300 mL). The pH of the aqueous layer was adjusted to 10by the slow addition of 5 N sodium hydroxide with vigorous stirring. Theorganic layer was removed using a separatory funnel and the aqueouslayer was extracted with dichloromethane (2×150 mL). The organics werecombined, dried over anhydrous sodium sulfate, and filtered. To thefiltrate was added diisopropylethylamine (2.83 mL, 16.25 mmol) anddi-tert-butyl dicarbonate (4.26 g, 19.5 mmol) and the resulting solutionwas stirred at room temperature overnight. The mixture was washed with1N hydrochloric acid, followed by a saturated solution of sodiumbicarbonate, and brine. The organic layer was dried over anhydroussodium sulfate, filtered, and evaporated under reduced pressure.Purification by MPLC (gradient eluant: 0-25% ethyl acetate/hexanes)afforded 1.58 g (34%) of the desired cis (R, S) isomer and 1.37 g (30%)of the undesired trans (S, S) isomer.

To a mixture the desired cis (R, S) product described in Step D,Intermediate 21 (1.51 g, 5.33 mmol) in a 1:1:1 solution oftetrahydrofuran/methanol/water (60 mL) was added solid LiOH (1.12 g,26.7 mmol) and the resulting solution was heated to 60° C. and stirredfor 18 h. The mixture was left standing to cool to room temperature andthen concentrated to remove the organic solvent. The aqueous layer wasacidified by the slow addition of 6 N hydrochloric acid to adjust the pHto 4 or 5. The acidic aqueous layer was extracted with dichloromethane(3×100 mL) and the organics were combined, dried over anhydrous sodiumsulfate, filtered, and evaporated under reduced pressure to affordIntermediate 21 (1.30 g, 91%) as a yellow oil. After two weeks standingat room temperature, the material solidified.

This intermediate was prepared in an analogous fashion to Intermediate19, except Intermediate 11 was replaced with Intermediate 21. LC-MS forC₁₈H₂₂F₃N₃ O calculated 353.17, found [M+H]⁺354.2.

This intermediate was prepared in an analogous fashion to Intermediate19, except Intermediate 11 was replaced with Intermediate 12. LC-MS forC₁₇H₁₉F₆N₃O calculated 395.17, found [M+H]⁺396.2.

This intermediate was prepared in an analogous fashion to Intermediate20, except Intermediate 11 was replaced with Intermediate 12. LC-MS forC₁₇ H₁₉ F₆ N₃ O₂calculated 411.17, found [M+H]⁺412.2.

This intermediate was prepared in an analogous fashion to Intermediate19, except Intermediate 11 was replaced with Intermediate 13. LC-MS forC₁₈H₂₁F₆N₃O calculated 409.17, found [M+H]⁺410.2.

To a stirred solution of phenyl magnesium bromide (3 M solution inether, 680 mL, 2 mol) in ethyl ether (500 mL) was addedexo-epoxynorbornane (150 g, 1.36 mol) in ethyl ether (250 mL) slowly.After the initial exotherm, the reaction was heated to reflux for 3 h,after which time it was cooled in an ice bath and quenched with water(25 mL). The resulting solution was diluted with ethyl ether and washedwith aqueous 3 N HCl twice. The combined aqueous layers where backextracted with ethyl ether twice and the combined organic layers wherewashed with brine, dried over MgSO₄, filtered, and concentrate underreduced pressure (100 mmHg, 30° C.) to give 230 g of a crude orange oil.This material was subject to flash chromatography (silica gel, 40% ethylether/hexanes) to give 67 g of pure product (45%). ¹H NMR (500 MHz,CDCl₃): □6.06 (d, J=1.0 Hz, 2H), 3.76 (s, 1H), 2.75 (d, J=2.0 Hz, 2H),1.86 (br s, 2H), 1.71-1.68 (m, 2H).

To a cooled (−78° C.) solution of oxalyl chloride (83 g, 660 mmol) indichloromethane (500 mL) was added DMSO (78 mL, 1.1 mol) indichloromethane (200 mL) rapidly but keeping the temperature below −50°C. To this solution was immediately added the product from Step A (67 g,610 mmol) in dichloromethane (600 mL) rapidly, but keeping thetemperature below −50° C. After stirring for 15 min at −78° C. thissolution was treated with triethylamine (310 mL, 2.1 mol) and allowed towarm to room temperature. After 1 h at room temperature, the reactionwas quenched with water and concentrated under reduced pressure. Thecrude residue was dissolved in a 3:1 solution of ethyl ether andpetroleum ether and washed 3 times with aqueous 1N HCl then with brine.The organic layer was dried over Na₂SO₄, filtered, and concentratedunder reduced pressure. The resulting residue was quicklychromatographed (short column—silica gel, 15% ethyl ether/hexanes) andconcentrated under reduced pressure. Final purification was achieved bydistillation (collecting the 60° C. to 70° C. fractions at 30 mm Hg) togive 18.5 g of pure product as a colorless liquid (28%). ¹H NMR (500MHz, CDCl₃): □ 6.53 (br s, 2H), 2.82 (br s, 2H), 1.97 (d, J=7.0 Hz, 2H),1.21 (dd, J=4.5, 6.5 Hz, 2H).

The product from Step B (17.5 g, 162 mmol) was combined withp-toluenesulfonic acid (4.9 g, 26 mmol) and ethylene glycol (13.1 mL,243 mmol) in benzene (200 mL) and heated to reflux. After 5 h, thesolution was allowed to cool to room temperature and stir overnight,after which time it was partitioned between ethyl ether and aqueoussaturated NaHCO₃. The organic phase was washed with brine, dried overMgSO₄, filtered and concentrated. The product was purified by flashchromatography (silica gel, 10% ethyl ether/hexanes) to give 19.0 g of acolorless oil (83%). ¹H NMR (500 MHz, CDCl₃): □6.18 (br s, 2H), 3.92 (t,J=6.0 Hz, 2H), 3.85 (t, J=6.0 Hz, 2H), 2.53 (br s, 2H), 1.92 (d, J=7.5Hz, 2H), 0.97 (dd, J=3.5, 10.5 Hz, 2H).

A solution of the product from Step C (2.0 g, 13 mmol) in a mixture ofmethanol (30 mL) and dichloromethane (24 mL) was cooled to −78° C. andtreated with ozone gas (7.5 psi, 2 L/min) until a blue tint to thesolution was apparent. At this time, the reaction was purged withnitrogen gas to remove the excess ozone and sodium borohydride (600 mg,16 mmol) was added to the reaction. The reaction was allowed to warm to0° C. on an ice bath before acetone was added to quench the excessreducing agent. The resulting solution was concentrated under reducedpressure and the product was purified by flash chromatography (silicagel, eluting with ethyl acetate) to give 1.9 g of a colorless oil whichupon cooling to −20° C. became a colorless solid (78%). ¹H NMR (500 MHz,CDCl₃): d 4.02 (m, 4H), 3.67 (m, 4H), 2.22 (t, J=6.0 Hz, 2H), 1.83 (m,2H), 1.63 (m, 2H).

To a cooled (−15° C.) solution of the product from Step D (1.26 g, 6.71mmol) in tetrahydrofuran (21 mL) was added n-butyllithium (2.5 M inhexanes, 2.8 mL, 7.0 mmol). After the reaction was stirred for 30 min at−15° C., tosyl chloride (1.28 g, 6.71 mmol) in tetrahydrofuran (10 mL)was added dropwise and the reaction was warmed to room temperature andstirred for an additional 30 min before being concentrated under reducedpressure. The mono-tosylate product was separated from small amounts ofstarting material and di-tosylation product by medium pressure liquidchromatography (silica gel, 40-100% ethyl acetate/hexanes) to give 900mg of a colorless oil (39%) which was used directly in the next step.

The product from Step E (707 mg, 2.07 mmol) was combined with sodiumhydride (60% dispersion in mineral oil, 250 mg) in tetrahydrofuran andstirred at room temperature. After 2 h the reaction was quenched withhydrogen chloride (2 N solution in ethyl ether, 4 mL) and the resultingprecipitate was filtered off. The filtrate was concentrated and purifiedby flash chromatography (silica gel, 20% ethyl ether/hexanes) to give320 mg of product (91%). ¹H NMR (500 MHz, CDCl₃): d 3.97 (m, 4H), 3.93(d, J=10.5 Hz, 2H), 3.57 (dd, J=2.5, 11.0 Hz, 2H), 1.84-1.81 (m, 2H),1.75 (m, 4H).

The product from Step F (250 mg, 1.47 mmol) was dissolved in a mixtureof tetrahydrofuran (4 mL) and aqueous 5% HCl (2 mL) and stirred at roomtemperature. After 18 h the reaction was diluted with ethyl ether,washed with brine, and dried over MgSO₄, filtered and concentrated underreduced pressure. The product was purified by flash chromatography(silica gel, 30% ethyl ether/hexanes) to give 51 mg of a volatile liquid(28%). ¹H NMR (500 MHz, CDCl₃): d 3.99 (dd, J 2.5, 11.0 Hz, 2H), 3.87(d, J=11 Hz, 2H), 2.28 (br s, 2H), 2.03 (m, 2H), 1.99 (m, 2H).

A solution of methyl-3-oxocyclopentane-carboxylate (20 g, 160 mmol) andtrimethyl orthoformate (85 mL, 780 mmol) in methanol was treated with acatalytic amount of p-toluenesulfonic acid. (3 g, 15.6 mmol) and theresulting solution was stirred for 4 h at room temperature. The solventwas evaporated under reduced pressure and the residue was then dissolvedin ether (600 mL). The solution was washed with saturated sodiumbicarbonate (2×200 mL), water (150 mL), brine (200 mL), dried overanhydrous sodium sulfate, filtered, and the solvent evaporated asbefore. Purification by flash column (eluant: 25% ether/pentane)afforded 21.52 g (73%) of the desired product as a clear oil. ¹H NMR(500 MHz, CDCl₃) δ 3.68 (s, 3H), 3.21 (d, J=9.9 Hz, 6H), 2.89 (p, J=8.5Hz, 1H), 2.14-2.05 (m, 2H), 2.02-1.80 (m, 4H).

A flame dried 500 mL round bottom flask was charged with 150 mL of drytetrahydrofuran, and then, set under nitrogen and cooled to −78° C.using an acetone/dry ice bath. Diisopropylamine (19.2 mL, 137 mmol) wasadded to the cooled solvent via syringe. 2.5 M n-butyllithium in hexanes(55 mL, 140 mmol) was slowly added to the solution. After 5 minstirring, the methyl ketal described in Step A, Intermediate 3 (21.52 g,114.4 mmol) in 50 mL of tetrahydrofuran was added dropwise via syringeand the resulting mixture was stirred at −78° C. for 2 h. 2-iodopropane(34.3 mL, 343 mmol) was then added dropwise via syringe and theresulting mixture was stirred overnight allowing it to warm slowly toroom temperature. The reaction was quenched with a solution of 10%citric acid and the organics were separated. The aqueous layer wasextracted with ether (3×150 mL) and all the organics were combined,dried over anhydrous sodium sulfate, filtered, and evaporated underreduced pressure. The crude product was purified by flash column usingan eluant of 20% ether/pentane to afford 16.74 g (64%) of the desiredproduct. ¹H NMR (400 MHz, CDCl₃) d 3.69 (s, 3H), 3.18 (d, J=20.5 Hz,6H), 2.57 (d, J=13.9 Hz, 1H), 2.29-2.20 (m, 1H), 1.90 (p, J=6.8 Hz, 1H),1.88-1.80 (m, 2H), 1.69-1.61 (m, 2H), 0.89 (dd, J=11.9 Hz, 6.8 Hz, 6H).

A solution of the ester from Step B, Intermediate 3 (16.74 g, 72.7 mmol)in ethanol (30 mL) was treated with 5 M aqueous NaOH (55 mL) and theresulting mixture was heated to reflux for 3 days. The mixture was thencooled to room temperature and acidified with concentrated hydrochloricacid. The organic solvent was evaporated under reduced pressure and theaqueous layer was then extracted with dichloromethane (5×100 mL). Theorganic extracts were combined, dried over anhydrous magnesium sulfate,filtered, and evaporated in vacuo to yield the crude 3-oxocyclopentanecarboxylic acid (11.07 g, 90%) as a yellow oil. Purification was notattempted because of the compounds polarity and lack of a chromophore.¹H NMR (500 MHz, CDCl₃) d 2.70 (d, J=18.1 Hz, 1H), 2.44-2.39 (m, 1H),2.30-2.15 (m, 2H), 2.14 (dd, J=18.1, 1.0 Hz, 1H), 2.06 (p, J=6.9 Hz,1H), 1.98 (m, 1H), 0.98 (dd, J=11.4, 6.9 Hz, 6H).

To a solution of the acid from Step C (540 mg, 3.20 mmol) indichloromethane (50 mL) was added oxalyl chloride (0.834 mL, 9.60 mmol)followed by 2 drops of N,N-dimethylformamide. The solution was stirredat room temperature for 80 min and then evaporated under reducedpressure. The residue was dissolved in dichloromethane (2 mL) and addedvia syringe to a prepared solution of Intermediate 2 (880 mg, 3.20 mmol)and triethylamine (0.820 mL, 6.50 mmol) in dichloromethane (20 mL). Theresulting mixture was stirred at room temperature for 18 h and thenquenched with water (25 mL). The organics were separated, washed withsaturated sodium bicarbonate and brine, dried over anhydrous sodiumsulfate, filtered, and evaporated. The crude product was purified byMPLC using a step-wise gradient eluant of 0-70% ethyl acetate/hexanes toafford Intermediate 2 (720 mg, 64%). ¹H NMR (500 MHz, CDCl₃).

Resolution of product from Step D, Intermediate 27 was accomplished bychiral separation using an HPLC equipped with a preparative ChiralPak ADcolumn. The separation was accomplished by injecting 100 mg/run andusing an eluant of 25% isopropanol and 75% heptane with a flow rate of 9mL/min.

To a solution of the pyridone from Step A, Intermediate 10 (7.50 g, 37.6mmol) in 200 mL of 2 M methanol/ammonia was added 1-benzoyl-4-piperdone(8.42 g, 41.4 mmol) and the mixture was heated at 60° C. for 18 h. Thesolvent was evaporated and the crude mixture was subject tochromatography, eluting with hexanes/ethyl acetate (50-70%). 10.2 g(96%) of the title product was collected. LC-MS for C₁₅H₁₃N₃O₃calculated 283.10 found 284.15 [M+H]⁺.

A mixture of the product from Step A (10.2 g, 36.0 mmol) and Pd/C (1.1g) in methanol (400 mL) was stirred overnight under a hydrogenatmosphere and then filtered through celite. Purification by columnchromatography eluting with hexanes/ethyl acetate (1:1) and methanol(5%) afforded 6.53 g (72%) of the title product. ¹H NMR (CD₃OD, 500 MHz)d7.98 (s, 1H), 7.46 (b, 6H), 6.83 (b, 2H), 4.84-4.44 (b, 2H), 3.72-3.67(b, 2H), 3.09-2.94 (b, 2H). LC-MS for C₁₅H₁₅N₃O calculated 253.12 found254.15 [M+H]⁺.

A mixture of the amine from step B (6.50 g, 25.6 mmol) and 30 mL of 20%sulfuric acid at 0° C. was treated with a solution of sodium nitrite(1.86 g, 28.2 mmol) in water (15 mL) via a syringe. After stirringvigorously at 0° C. for 25 min a small crystal of urea was added. Theresulting deep red mixture was added slowly, via a cannula, to 150 mL of20% sulfuric acid at 90° C. The flask was removed from the oil bathimmediately upon completion of addition (10 min) and the mixture wascooled to room temperature. The pH was adjusted to 7 with potassiumcarbonate and the resulting precipitate was filtered off. The filtratewas the extracted with dichloromethane and the combined organic layerswere washed with brine, dried (MgSO₄) and concentrated in vacuo.Purification by column chromatography eluting with hexanes/ethyl acetate(1:1) and 4% methanol afforded 4.68 g (72%) of the title product. LC-MSfor C₁₅H₁₄N₂O₂ calculated 254.11 found 255.1 [M+H]⁺. The aqueous layercontained the deprotected amine.

A flame dried, 3-neck round bottom flask containing a suspension of 0.71g (18 mmol) sodium hydride (60% dispersion in mineral oil) and anhydrousN,N-dimethylformamide (30 mL) under N₂ was stirred for 10 min. Theproduct from Step C (3.0 g, 12 mmol) in N,N-dimethylformamide (30 mL)was then added slowly via a cannula and the resulting creamish brownmixture was stirred at room temperature for 45 min then at 50° C. for 30min. After cooling to room temperature carbon disulfide (3.5 mL, 59mmol) was added slowly and the resulting dark brown mixture was stirredat room temperature for 2 h. Iodomethane (3.07 mL, 47.2 mmol) was thenadded slowly via a syringe and after stirring for 30 min, the reactionwas quenched with water. The suspension was diluted with ethyl acetate,extracted and the combined organic layers were dried (MgSO₄) andconcentrated in vacuo. The resulting brown oil was chromatographedeluting with hexanes/ethyl acetate (40-60%) to afford 3.52 g (87%) ofthe title product. LC-MS for C₁₇H₁₆N₂O₂S₂ calculated 344.07 found 345.1[M+H]⁺.

A flame dried, 3-neck 500 mL round bottom flask containing a suspensionof 13.8 g (45.9 mmol) 1,3-dibromo-5,5-dimethylhydantoin indichloromethane (200 mL) was stirred at room temperature for 10 min andthen cold to −78° C. 100 g (80 eq) of hydrogen fluoride/pyridine (70%)solution was then added slowly via a syringe and the resulting clearsolution was stirred at −78° C. for 30 min. 3.5 g, (10.2 mmol) of theproduct from Step C in dichloromethane (60 mL) was then added via acannula and the resulting creamish/yellow mixture was stirred at −5° C.for 2 h. The mixture was diluted with ether at −5 ° C. and quenched witha cold solution of sodium bicarbonate and sodium bisulfate until the redcolor disappeared. The pH was adjusted to 7-8 with 5.0 N NaOH and thelayers were separated. The organic layer was dried (MgSO₄) andconcentrated in vacuo. Flash chromatography, eluting with hexanes/ethylacetate (40-50%) afforded 2.47 g (75%) of the title product. LC-MS forC₁₆H₁₃F₃N₂O₂ calculated 322.09 found 323.2 [M+H]⁺.

A solution of the product from Step E (2.45 g, 7.60 mmol) in 20 mLconcentrated HCl was stirred at 75° C. for 18 h and concentrated. Theresulting oil was dissolved in dichloromethane (200 mL) and stirred withCa(OH)₂ (2.0 g) for 30 min. The white precipitate was filtered throughcelite, and the filtrate was concentrate to afford 1.52 g (92%) of theproduct, Intermediate 28. ¹H NMR (CD₃OD, 500 MHz) d8.34 (s, 1H), 7.22(s, 1H), 4.04 (s, 2H), 3.25-3.22 (t, 2H), 2.97-2.95 (t, 2H). LC-MS forC₉H₉F₃N₂O calculated 218.07 found 219.05 [M+H]⁺.

The procedure described in Step A, Intermediate 19 were followed butusing intermediate 28 instead of intermediate 8. LC-MS for C₂₃H₃₂F₃N₃O₄calculated 471.23 found 372.25 [M+H−Boc]⁺.

A solution of the product from Step A in ethyl acetate at 0° C. wastreated with a saturated solution of HCl in ethyl acetate and themixture was stirred for 2 h. The volatiles were evaporated in vacuo toafford a white foam, Intermediate 29 LC-MS for C₁₈H₂₄F₃N₃O₂ calculated371.18 found 372.25 [M+H]⁺.

A mixture of ethyl 2,2-dimethyl-methylacetoacetate (3.0 g, 21 mmol),ethylene glycol (3.8 g, 62 mmol), camphorsulfonic acid (50 mg) andbenzene (50 mL) was refluxed in a Dean-Stark apparatus, with continuesremoval of water. After ensuring the completion of the reaction (by TLC)it was diluted with water and extracted with ether (100 mL). The etherlayer was washed with brine, dried (anhydrous magnesium sulfate) andconcentrated to afford the desired compound (4.1 g). This was taken inether (50 mL) and was slowly added to lithium aluminum hydride (1.2 g,32 mmol) at 0° C. The reaction was warmed to room temperature andstirred for 12 h. The reaction mixture was then quenched sequentiallywith water (1.5mL), 15% NaOH (1.5 mL) and water (4.5 mL). The resultantheterogeneous mixture was vigorously stirred and filtered. Evaporationof the filtrate followed by flash column chromatography eluting withhexanes/ethyl acetate (4:1) gave 2.2 g of the title compound.

To a stirring slurry of silica (12 g, 230-400 mesh) in methylenechloride (100 mL) was added a 10% aqueous solution of oxalic acidfollowed by the product from step A (2.0 g, 13 mmol) in methylenechloride (5 mL). The resultant mixture was stirred at room temperatureuntil the reaction was complete. Upon the completion of the reaction,NaHCO₃ (1.0 g) was added. The reaction was stirred for 10 min and thenfiltered. The filtrate was evaporated to give the 1.5 g of the titlecompound that required no purification.

To a premixed solution of triethyl orthoformate (1.3 g, 8.6 mmol), tin(IV) chloride (8.6 mL 1.0 M solution in dichloromethane, 8.6 mmol) at−40° C. was added the ketone from Step B (0.5 g, 4.3 mmol) indichloromethane (3 mL). The reaction mixture was warmed to −5° C. over1.5 h before being quenched with saturated NaHCO₃ solution and extractedwith ether (2×50 mL). The ether layer was washed with brine, dried(anhydrous magnesium sulfate), concentrated and purified by flash columnchromatography. Eluting with hexanes/ether (9:1) gave the title compound(0.23 g, 43%).

The intermediate from Step C (0.23 g) in hexanes (5 mL) and Pd/C (5%, 10mg) was hydrogenated at room temperature using a hydrogen filled balloonuntil TLC indicated the completion of reaction. The reaction mixture wasfiltered and the filtrate was carefully evaporated (volatile product!)to yield the mixtures of the desired Intermediate 30 and the overreduction product. The recovery of Intermediate 30 was furtherfacilitated by a subsequent TPAP/NMMO/dichloromethane oxidation of themixture, which after 1 h was filtered to yield 221 mg of the titlecompound that required no further purification. ¹H NMR (CDCl₃, 500 MHz):d 3.98 (t, 2H), 3.58 (s, 2H), 2.56 (t, 2H), 1.15 (s, 6H).

Following Steps A-D given for the preparation of intermediate 30 andstarting from methyl 2,4-dimethyl-3-oxobutyrate, gave the titlecompound. ¹H NMR (CDCl₃, 500 MHz): d 4.22 (m, 1H), 3.99 (m, 1H), 3.62(m, 1H), 3.28 (m, 1H), 2.72 (m, 1H), 1.16 (d, J=6.8 Hz, 3H), 0.97 (d,J=6.8 Hz, 3H).

Prepared according to J. Am. Chem. Soc., 1997, 119, 4285, except thatthe reaction was performed on the ethyl ester.

To a solution of t-butyl 4-oxo-1-piperidinecarboxylate (5.0 g, 25 mmol)in tetrahydrofuran (50 mL) at −10° C. was added a solution of lithiumbis(trimethylsilyl)amide (25 mmol, 1 M solution in tetrahydrofuran) andthe resultant solution was stirred for 1 h while the temperature wasraised to 0° C. 2-chloro-1,3-bis(dimethylamino)trimethiniumhexafluorophosphate (11.5 gm, 37.6 mmol) was added in one lot and thestirring was continued for an additional 20 min at 0° C. and then atroom temperature for 2 h. Ammonium acetate (4.83 gm, 63.0 mmol) wasadded to the above and the resultant reddish brown mixture was stirredfor 4 h at 60° C. The reaction mixture was cooled and extracted withether (2×100 mL). The organic layer was dried (MgSO₄) and concentratedin vacuo. Flash chromatography eluting with hexanes/ethyl acetate(10-20%) afforded 3.97 g (60%) of the title product.

¹H NMR (CDCl₃, 500 MHz): d 8.39 (s, 1H), 7.43 (s, 1H), 4.59 (s, 2H),3.75 (t, J=5.7 Hz, 2H), 2.98 (t, J=5.7 Hz, 2H), 1.50 (s, 9H).

A 3-neck, flame dried, round bottom flask containing Intermediate 33(500 mg, 1.86 mmol), Iron (III) acetylacetonate (0.032 g, 0.090 mmol)and 10 mL tetrahydrofuran/N-methyl-2 -pyrrolidone (9:1) at 0° C. wastreated with 2.21 mL (1.0 M) of cyclohexyl magnesium bromide. Theorange/red color immediately disappear and the resulting dark brownmixture was stirred over the weekend. The reaction was quenched withsaturated aqueous ammonium hydroxide and extracted with ether. Flashchromatography eluting with hexanes/ethyl acetate (15%) afforded 0.295 gof the title product. ¹H NMR (CD₃ OD, 500 MHz) d8.29 (s, 1H), 7.24 (s,1H), 4.59 (s, 2H), 3.78-3.74 (t, 2H), 2.98 (t, 2H), 2.52 (b, 1H),1.88-1.86 (b, 2H), 1.79-1.77 (b, 1H), 1.51-1.49 (b, 13H), 1.45-1.40 (t,2H), 1.3-1.28 (b, 1H). LC-MS for C₁₉H₂₈N₂O₂ calculated 316.22 found317.15 [M+H]⁺.

A solution of the product from Step A in ethyl acetate at 0° C. wastreated with a saturated solution of HCl in ethyl acetate and theresulting mixture was stirred for 2 h. The volatiles were evaporated invacuo to afford a white foam, Intermediate 34. LC-MS for C₁₄H₂₀N₃calculated 216.32 found 217.32 [M+H]⁺.

Starting from Intermediate 33 (0.8 g, 3 mmol) and cyclopentylmagnesiumbromide (1.5 mL, 2 M solution in ether) using a procedure analogous tointermediate 34, Step A yielded 0.245 g of the title compound. ¹H NMR(CD₃ OD, 500 MHz) d8.32 (s, 1H), 7.27 (s, 1H), 4.58 (s, 2H), 3.75 (t,2H), 2.98 (t, 2H), 2.51-1.58 (m, 11H), 1.51 (s, 9H).

A solution of the product from Step A in ethyl acetate at 0° C. wastreated with a saturated solution of HCl in ethyl acetate and theresulting mixture was stirred for 2 h. The volatiles were evaporated invacuo to afford 0.230 g of Intermediate 35. LC-MS for C₁₃H₁₈N₃calculated 202.15, found 203.4 [M+H]⁺.

This intermediate was synthesized in a series of steps analogous tothose described for Intermediate 16, except that in Step A acetaldehydewas replaced with ethyl 2-bromopropionate.

A solution of intermediate 19 (890 mg, 2.08 mmol),tetrahydro-4H-pyran-4-one (320 mg, 3.13 mmol), diisopropylethylamine(1.10 mL, 6.24 mmol) and crushed molecular sieves (4 Å, 500 mg) indichloromethane (50 mL) was treated with sodium triacetoxyborohydride(2.20 g, 10.4 mmol) and stirred at room temperature overnight. Thereaction was quenched with saturated sodium bicarbonate solution (50 mL)and diluted with an additional 25 mL of dichloromethane. The organiclayer was separated and the aqueous layer was washed withdichloromethane (2×25 mL). The organics were combined, dried overanhydrous sodium sulfate, filtered and evaporated under reducedpressure. The crude product was purified by reverse phase HPLC to yieldExample 1 (915 mg, 86.0%). LC-MS for C₂₃H₃₁F₃N₃O₂ calculated 439.24,found [M+H]⁺440.2.

EXAMPLE 2

To a solution of product described in Example 1 (136 mg 0.265 mmol) andcrushed 4Å molecular sieves (100 mg) in dichloromethane (20 mL) wasadded formalin (0.2 mL) and the resulting suspension was stirred for 30min at room temperature. This mixture was then treated with sodiumtriacetoxyborohydride (280 mg, 1.33 mmol) and stirred an addition 15 hat room temperature. The reaction was quenched with saturated sodiumbicarbonate solution (20 mL) and diluted with an additional 10 mL ofdichloromethane. The organic layer was separated and the aqueous layerwas extracted with dichloromethane (2×20 mL). The organics werecombined, dried over anhydrous sodium sulfate, filtered and evaporatedunder reduced pressure. The crude product was purified by reverse phaseHPLC to yield Example 5 (80 mg, 57.6%). LC-MS for C₂₄H₃₅F₃N₃O₂calculated 453.26, found [M+H]⁺454.3.

EXAMPLE 3

Example 3 was prepared as detailed in Example 2 using acetaldehydeinstead of formaldehyde. LC-MS for C₂₅H₃₇F₃N₃O₂[M⁺H⁺] calculated 468.28,found 468.25.

EXAMPLE 4

Example 4 was prepared as detailed in Example 2 usingbenzyloxyacetaldehyde instead of formaldehyde. LC-MS forC₃₂H₄₃F₃N₃O₂[M⁺H⁺] calculated 574.32, found 574.35.

EXAMPLE 5

A mixture of Example 4 (43 mg, 0.075 mmol), 10% Pd/C (10 mg), andethanol (5 mL) was stirred at room temperature under a hydrogen balloonfor 18 h before being filtered and concentrated to dryness. The crudeproduct was purified by reverse phase HPLC to yield Example 5 (13.3 mg,36.9%). LC-MS for C₂₅H₃₇F₃N₃O₃[M⁺H⁺] calculated 484.27, found 484.3.

EXAMPLE 6

To Intermediate 10 (0.25 g, 1 mmol) and K₂CO₃ (0.5 g, 3.6 mmol) in dryN,N-dimethylformamide (5.0 mL) at 75° C. was bubbled CHClF₂ through thereaction vessel attached to a cold finger (at −78° C.) for 20 min. Themixture was stirred for an additional 2 h at 75° C., then allowed tocool to room temperature and stir overnight. The reaction mixture wasdiluted with water and extracted with ethyl acetate (2×25 mL). Thesolvent layer was washed with brine, dried (MgSO₄), and concentrated invacuo. Purification was carried out by flash column chromatography(eluant: 95% hexanes/ethyl acetate) to afford 0.06 g (20%) of the titleproduct. ¹H NMR (500 MHz, CDCl₃): 8.32 (d, J=2.1 Hz, 1H), 7.25 (d, J=2.1Hz, 1H), 6.54 (t, J=72.7 Hz, 1H), 4.62 (s, 2 H), 3.77 (t, J=6.0 Hz, 2H),3.01 (t, J=5.7 Hz, 2H), 1.52 (s, 9H).

To a solution of the intermediate from Step A (0.06 g) in ethyl acetate(1.0 mL) was added a solution of HCl in ethyl acetate. The resultingsolution was stirred for 30 min. Volatiles were removed under vacuum togive the desired product (0.054 g) as the HCl salt. LC-MS for C₉H₁₀F₂N₂Ocalculated 200.19, found [M+H]⁺201.05.

A mixture of Intermediate 11 (107 mg, 0.400 mmol), the intermediate fromStep B, Example 6 (0.053 g, 0.27 mmol), 4-dimethylaminopyridine (2 mg)and bromo-tris-pyrrolidino-phosphonium hexafluorophosphate (0.247 g,0.53 mmol) in dichloromethane (5.0 mL) was treated withdiisopropylethylamine (0.27 mL, 1.6 mmol) and the resulting mixture wasstirred overnight. The reaction was quenched with water and extractedwith ethyl acetate. The organic layer was washed with brine, dried(anhydrous MgSO₄)) and concentrated in vacuo. Purification was carriedout by preparative TLC (eluant: hexanes/ethyl acetate (1:1)) to afford65.5 mg (55%) of the title product. LC-MS for C₂₃H₃₃F₂N₃O₄ calculated453.25 found [M+H]⁺454.2.

To a solution of the intermediate from Step D (0.065 g) in ethyl acetate(2.0 mL) was added a solution of HCl in ethyl acetate. The resultingsolution was stirred for 30 min. Removal of the volatiles under reducedpressure gave the desired product (0.06 g) as the HCl salt. LC-MS forC₁₈H₂₅F₂N₃O₂ calculated 353.20, found [M+H]⁺354.2.

A solution of the intermediate from Step D (0.025 g, 0.070 mmol) indichloromethane (2.5 mL) and diisopropylethylamine (0.042 mL) wastreated with tetrahydro-4H-pyran 4-one (0.035 g, 0.35 mmol) and 4 Åmolecular sieve. After stirring the mixture for 45 min, sodiumtriacetoxyborohydride (0.074 g, 0.35 mmol) was added. After 18 h themixture was filtered and the filtrate was concentrated in vacuo.Reverse phase HPLC purification of the crude gave the title product,which was subsequently transformed to the HCl salt (0.018 g). LC-MS forC₂₃H₃₃F₂N₃O₃ calculated 437.26, found [M+H]⁺438.25.

EXAMPLE 7

To a solution Intermediate 10 (0.098 g, 0.40 mmol) in dichloromethane(4.0 mL) was added p-fluorophenyl boronic acid (0.112 g, 0.800 mmol),copper (II) acetate (160 mg, 0.800 mmol), triethylamine (0.54 mL, 2.0mmol) and 4 Å (500 mg) molecular sieves. The resultant mixture wasstirred 48 h and filtered. The filtrate was concentrated and purified bypreparative TLC (eluant: 1:1 hexanes/ethyl acetate) to afford 0.05 g ofthe N-Boc intermediate. This intermediate was subsequently transformedto the title compound by treating with 20% sulfuric acid to afford 0.094g of the product. LC-MS for C₁₄H₁₃FN₂O calculated 244.11, found[M+H]⁺245.15.

A mixture of Intermediate 11 (0.11 g, 0.41 mmol), the intermediate fromStep A, Example 7 (0.09 g, 0.2 mmol), 4-dimethylaminopyridine (2 mg) andbromo-tris-pyrrolidino-phosphonium hexafluorophosphate (0.233 g, 0.5mmol) in dichloromethane (5.0 mL) was treated with diisopropylethylamine(0.21 mL, 1.2 mmol) and the mixture was stirred overnight. The reactionwas quenched with water and extracted with ethyl acetate. The organiclayer was washed with brine, dried (anhydrous MgSO₄)) and evaporated invacuo. Purification was carried out by preparative TLC (eluant:hexanes/ethyl acetate (1:1)) to afford 14.7 mg (32%) of the titleproduct. LC-MS for C₂₈H₃₆FN₃O₄ calculated 497.28 found [M+H]⁺498.4.

To a solution of the intermediate from Step B, Example 7 (0.014 g) inethyl acetate (1.0 mL) was added a solution of HCl in ethyl acetate. Theresulting solution was stirred for 30 min. Removal of the volatilesunder reduced pressure gave the desired product (0.013 g) as the HClsalt. LC-MS for C₂₃H₂₈FN₃O₂ calculated 397.22, found [M+H]⁺398.4.

A solution of the intermediate from Step C, Example 7 (0.013 g, 0.070mmol) in dichloromethane (2.0 mL) and diisopropylethylamine (0.042 mL)was treated with tetrahydro-4H-pyran 4-one (0.035 g, 0.35 mmol) and 4 Åmolecular sieve. After stirring the mixture for 45 min, sodiumtriacetoxyborohydride (0.074 g, 0.35 mmol) was added. After 18 h, themixture was filtered and the filtrate was evaporated in vacuo. Reversephase HPLC purification of the crude afforded the title product, whichwas subsequently transformed to the HCl salt (0.007 g). LC-MS forC₂₈H₃₆FN₃O₃ calculated 481.28, found [M+H]⁺482.2.

EXAMPLE 8

To a solution of Intermediate 10 (100 mg, 0.40 mmol) inhexamethylphosphoramide (3.0 mL) at room temperature was added asolution of sodium hydroxide (32 mg, 0.80 mmol) in water (0.5 mL). Afterstirring for 5 min, 2-Iodopropane (0.079 mL, 0.80 mmol) was added to themixture and the resultant reddish brown mixture was stirred overnight.The mixture was extracted with ethyl acetate and washed with water,brine, dried (MgSO₄) and concentrated in vacuo. Column chromatographyeluting with hexanes/ethyl acetate (20-30%) afforded 0.075 g (70%) ofthe title product. ¹H NMR (500 MHz, CDCl₃):8.120 (s, 1H), 6.92 (s, 1H),4.57 (s, 2H), 4.54 (m, J=2.4 Hz, 1H), 3.74 (t, 2H), 2.94 (t, 2H), 1.50(s, 9H), 1.36 (d, J=2.4 Hz, 6H).

To the intermediate from Step A, Example 8 (75 mg, 0.28 mmol) was addeda solution 4 N HCl in dioxane (2.0 mL) and the resulting mixture wasstirred for 30 min. Evaporation of the volatiles in vacuo, afforded0.070 g of the title compound. LC-MS for C₁₁H₁₆N₂O calculated 192.13,found [M+H]⁺193.1.

Starting from the intermediate prepared in Step B, Example 8 (0.070 g,0.26 mmol) and Intermediate 11 (80 mg, 0.31 mmol) and following theprocedure described in Step C, Example 3 gave 0.087 g of the titlecompound. LC-MS for C₂₅H₃₉N₃O₄ calculated 445.29, found [M+H]⁺446.3.

To a solution of the product described in Step C, Example 8 (0.087 g) inethyl acetate (2.0 mL) was added a saturated solution of HCl in ethylacetate (2.0 mL). the resulting solution was stirred for 30 min.Evaporation of the volatiles in vacuo, afforded 0.08 g of the amine HClsalt. LC-MS for C₂₀H₃₁N₃O₂ calculated 345.24, found [M+H]⁺346.25

A solution of the product described in Step D, Example 8 (0.040 g, 0.095mmol) in dichloromethane (2.0 mL) and diisopropylethylamine (0.042 mL)was treated with tetrahydro-4H-pyran 4-one (0.013 mL, 0.14 mmol) and 4 Åmolecular sieve. After stirring the mixture for 45 min, excess sodiumtriacetoxyborohydride was added. Stirring was continued for another 18h. The mixture was filtered and the filtrate was concentrated in vacuo.Reverse phase purification of the crude afforded 27.4 mg of the titleproduct. LC-MS for C₂₅H₃₉N₃O₃ calculated 429.30, found [M+H]⁺430.3.

EXAMPLE 9

To a solution of Intermediate 10 (0.1 g, 0.399 mmol) inhexamethylphosphoramide (3.0 mL) at room temperature was added 0.048 gsodium hydride (60% dispersion in mineral oil) and the resultingbrownish red mixture was stirred for 5 min. 2-Iodo-1,1,1-trifluoroethane (0.12 mL) was then added to the mixture via a syringe and themixture was stirred overnight. The reaction was quenched with water,extracted with ethyl acetate, dried (MgSO₄) and concentrated in vacuo.Column chromatography eluting with hexanes/ethyl acetate (20-30%)afforded 0.015 g (16%) of the title product ¹H NMR (CDCl₃, 500 MHz)d8.20 (s, 1H), 7.02 (s, 1H), 4.59 (s, 2H), 4.39 (q, 2H), 3.75 (t, J=3.5Hz, 2H), 2.97 (t, J=3.5 Hz, 2H), 1.50 (s, 9H).

To the intermediate from Step A, Example 9 (0.015 g) was added asaturated solution of HCl in ethyl acetate (2.0 mL). The resultingmixture was stirred for 30 min. Evaporation of the volatiles in vacuo,afforded 0.015 g of the title compound. LC-MS for C₁₀H₁₁F₃N₂O calculated232.08, found [M+H]⁺ 233.2.

Starting from the intermediate prepared in Step B, Example 9 (0.015 g,0.049 mmol) with Intermediate 11 (0.020 g, 0.073 mmol) and following theprocedure described in Step C, Example 3 gave 0.013 g of the titlecompound. LC-MS for C₂₄H₃₄F₃N₃O₄ [M+H]⁺ calculated 486.25, found 386.2[M+H−100(Boc)].

To a solution of the product described in Step C, Example 9 (0.013 g) inethyl acetate (2.0 mL) was added a 4 N solution of dioxane/HCl (2.0 mL).the resulting solution was stirred for 30 min. Evaporation of thevolatiles in vacuo, afforded 0.015 g of the amine HCl salt. LC-MS forC₁₉H₂₆F₃N₃O₂ calculated 385.20, found [M+H]⁺ 386.2.

To a solution of the intermediate from Step D, Example 9 (0.015 g, 0.033mmol) in dichloromethane (2.0 mL) and diisopropylethylamine (0.013 mL)was treated with tetrahydro-4H-pyran 4-one (0.006 mL) and 4 Å molecularsieve. The mixture was stirred for 45 min and sodiumtriacetoxyborohydride was added. After 18 h, the mixture was filteredand the filtrate concentrated in vacuo. Reverse phase purificationafforded 4.2 mg of the title product. LC-MS for C₂₄H₃₄F₃N₃O₃ calculated469.26, found [M+H]⁺ 470.2.

EXAMPLE 10

To a solution of the intermediate from Step D, Example 6 (0.035 g, 0.086mmol) in dichloromethane (2.0 mL) and diisopropylethylamine (0.038 mL)was treated with 3-methoxy-pyran-4-one (0.056 g) and 4 Å molecularsieve. The mixture was stirred for 45 min and sodiumtriacetoxyborohydride was added. After 18 h, the mixture was filteredand the filtrate was concentrated in vacuo. Reverse phase purification(ChiralCel OD column) afforded 4.7 mg of the less polar and 7.2 mg ofthe more polar title products. LC-MS for (less polar isomer)C₂₄H₃₅F₃N₃O₄ calculated 467.26, found [M+H]⁺ 468.25. LC-MS for (morepolar isomer) C₂₄H₃₅F₃N₃O₄ calculated 467.26, found [M+H]⁺ 468.25.

EXAMPLE 11

To a solution of Intermediate 10 (100 mg, 0.399 mmol) inhexamethylphosphoramide (3.0 mL) at room temperature was added 0.024 gsodium hydride (60% dispersion in mineral oil). The resulting brownishred mixture was stirred for 5 min. Bromocyclobutane (0.162 g) inhexamethylphosphoramide was then added to the mixture via a syringe andthe mixture was stirred overnight. The reaction was quenched with water,extracted with ethyl acetate, dried (MgSO₄) and concentrated in vacuo.Column chromatography eluting with hexanes/ethyl acetate (20-30%)afforded 0.007 g (6%) of the title product. ¹H NMR (400 MHz, CDCl₃)d8.06 (s, 1H), 6.86 (s, 1H), 4.64-4.67 (m, 1H), 4.57 (s, 2H), 3.72-3.75(t, J=3.75 Hz, 2H), 2.94-2.96 (t, J=3.75 Hz, 2H), 2.44-2.50 (m, 2H),2.15-2.21 (m, 2H), 1.88-1.91 (m, 1H), 1.70-1.76 (m, 1H), 1.51 (s, 9H).

To a solution of the intermediate from Step A, Example 11 (0.007 g) wasadded a saturated solution of HCl in ethyl acetate (2.0 mL). Theresulting solution was stirred for 30 min. Evaporation of the volatilesin vacuo, afforded 0.007 g of the title compound. LC-MS for C₁₂H₁₆N₂Ocalculated 204.13, found [M+H]⁺ 205.1.

Starting from the product from Step B, Example 11 (0.007 g, 0.03 mmol)with Intermediate 11 (0.010 g, 0.035 mmol) and following the proceduredescribed in Step C, Example 3 gave 0.005 g of the title compound. LC-MSfor C₂₆H₃₉N₃O₄ calculated 457.25, found 358.2 [M+H−100(Boc)].

To a solution of product described in Step C, Example 11 (0.005 g) inethyl acetate (2.0 mL) was added a 4 N solution of HCl in dioxane (2.0mL). The resulting solution was stirred for 30 min. Evaporation of thevolatiles in vacuo, afforded 0.004 g of the amine HCl salt. LC-MS forC₂₁H₃₁N₃O₂ calculated 357.20, found [M+H]⁺ 358.2.

To a solution of the intermediate from Step D, Example 11 (0.004 g,0.011 mmol) in dichloromethane (2.0 mL) and diisopropylethylamine (0.004mL) was added tetrahydro-4H-pyran 4-one (0.003 mL) and 4 Å molecularsieve. The mixture was stirred for 45 min and sodiumtriacetoxyborohydride was added. After 18 h, the mixture was filteredand the filtrate was concentrated. in vacuo. Reverse phase purificationafforded 4.2 mg of the title product. LC-MS for C₂₆H₃₉N₃O₃ calculated441.30, found [M+H]⁺ 442.3.

EXAMPLE 12

A solution of Intermediate 29 (50 mg, 0.11 mmol) in dichloromethane (2.0mL) and diisopropylethylamine (0.042 mL, 0.24 mmol) at room temperatureunder N₂ was treated with activated 4 Å molecular sieve (10 mg) andtetrahydro-4H-pyran-4-one (0.015 mL, 0.16 mmol). The mixture was stirredfor 45 min and sodium triacetoxyborohydride (69 mg, 0.33 mmol) wasadded. After stirring for 16 h, the reaction was quenched with saturatedaqueous sodium bicarbonate and filtered through celite. The layers wereseparated and the organic layer dried (MgSO₄), filtered andconcentrated. Reverse phase HPLC purification afforded 20.1 mg ofExample 12. LC-MS for C₂₃H₃₂F₃N₃O₃ calculated: 455.24 found: 456.25[M+H]⁺.

EXAMPLE 13

The product from Step A, Intermediate 29 (83 mg, 0.18 mmol), indichloromethane (1.5 mL) was treated with 3-chloroperoxybenzoic acid(163 mg, 0.700 mmol). The mixture was stirred for 2 h and then excessCa(OH)₂ was added and the reaction mixture was stirred for an additionalfor 30 min. The white precipitate was filtered through celite and thefiltrate was concentrated in vacuo to afford the title product as awhite foam. LC-MS for C₂₃H₃₂F₃N₂O₅ calculated: 487.23, found: 388.25[M+H−Boc]⁺.

A solution of the product from Step A in ethyl acetate at 0° C. wastreated with a saturated solution of HCl in ethyl acetate. The resultingsolution was stirred for 2 h. The volatiles were evaporated in vacuo toafford the title product as a white foam which was used without furtherpurification. LC-MS for C₁₈H₂₄F₃N₃O₃ calculated: 387.18, found 388.3[M+H]⁺.

Following the procedure described for Example 12 but using the productfrom Step B instead of intermediate 29 afforded 33 mg of Example 13.LC-MS for C₂₃H₃₂F₃N₃O₄ calculated: 471.23, found: 472.3 [M+H]⁺.

EXAMPLE 14

Following the procedure described for Example 12 but using Intermediate1 instead of tetrahydro-4H-pyran-4-one afforded Example 14 as mixture of4 diastereomers. Chiral separation on an OD column eluting withethanol/heptane (5%) afforded the 4 resolved diastereomers. LC-MS forC₂₄H₃₄F₃N₃O₃ calculated: 470.26 found: 470.15 [M+H]⁺.

EXAMPLE 15

A solution of the ester from Step C intermediate 16 (diastereomericmixture) (0.50 g, 1.3 mmol) in 5.0 mL of pyridine at 0° C. was treatedwith a catalytic amount of N,N-dimethyl-4-aminopyridine and 0.26 mL(2.75 mmol) of acetic anhydride. The resulting mixture was stirredovernight. The volatiles were evaporated and the product was purified byflash chromatography (eluting with hexanes/ethyl acetate 7:3). 0.527 gof the title product (mixture of 2 diastereomers) was collected. LC-MSfor C₂₂H₃₁NO₆ calculated: 405.22, found: 406.2 [M+H]⁺.

This acid was prepared following the procedure described in Step CIntermediate 12, and was used in the next step without furtherpurification. LC-MS for C₁₅H₂₅NO₆ calculated 315.17 found 338.2 [M+Na].

The procedure described in Step A, Intermediate 19 was followed, usingIntermediate 28 instead of Intermediate 8 and the acid from Step Babove. LC-MS for C₂₄H₃₂F₃N₃O₆ calculated: 515.22, found: 416.3[M+H−Boc]⁺.

A solution of the product from Step C in ethyl acetate at 0° C. wastreated with a saturated solution of HCl in ethyl acetate. The resultingsolution was stirred for 2 h. The volatiles were evaporated in vacuo toafford a white foam and used with out further purification. LC-MS forC₁₉H₂₄F₃N₃O₃ calculated: 415.17, found: 416.4 [M+H]⁺.

Following the procedure described for Example 12 but using the aminefrom Step D instead of intermediate 29 afforded the acetate protectedproduct. Hydrolysis of the acetate was accomplished by stirring in amixture of methanol/K₂CO₃(10 eq) at 70° C. Chiral separation on an ODcolumn eluting with ethanol/hexanes (7%) afforded the desired resolveddiastereomers (Example 15). LC-MS for C₂₂H₃₀F₃N₃O ₄ calculated: 457.22,found: 458.15 [M+H]⁺.

EXAMPLE 16

A solution of Example 15 (60 mg, 0.11 mmol) in methanol (2.0 mL) wastreated with 0.169 mL of formaldehyde (37%, 1.13 mmol) and sodiumcyanoborohydride (21 mg, 0.34 mmol). The mixture was stirred overnightand the volatiles were evaporated. The resulting oil was dissolved indichloromethane and washed with a small amount of water. The aqueouslayer was extracted with dichloromethane (×2) and the combined organiclayers were dried (MgSO₄) and concentrated in vacuo. Purification byreverse phase HPLC afforded 36.3 mg of Example 16. LC-MS forC₂₃H₃₂F₃N₃O₄ calculated: 471.23 found: 472.25 [M+H]⁺.

EXAMPLE 17

A solution of intermediate 19 (304 mg, 0.712 mmol), Intermediate 1 (160mg, 1.42 mmol), diisopropylethylamine (370 μL, 2.14 mmol) and crushedmolecular sieves (4 Å, 150 mg) in dichloromethane (25 mL) was treatedwith sodium triacetoxyborohydride (755 mg, 3.56 mmol) and stirred atroom temperature overnight. The reaction was quenched with saturatedsodium bicarbonate solution (25 mL) and diluted with an additional 25 mLof dichloromethane. The organic layer was separated and the aqueouslayer was washed with dichloromethane (2×20 mL). The organics werecombined, dried over anhydrous sodium sulfate, filtered and evaporatedunder reduced pressure. The residue was purified by preparative TLC(eluant: 0.5% NH₄OH/5% methanol/94.5% CH₂Cl₂) to yield 239 mg (74%) ofthe final product as a mixture of four diastereomers. Cis and transracemate in reference to the pyran ring were resolved by HPLC equippedwith a Preparative ChiralCel OD column (eluant: 5% ethanol/95% hexanes).Cis racemate was further resolved by using the Preparative ChiralPak ADcolumn (eluant: 5% ethanol/95% hexanes). LC-MS for C₂₄H₃₅F₃N₃O₂calculated 453.26, found [M+H]⁺ 454.3.

EXAMPLE 18

To a solution of a single isomer described in Example 17 (40 mg, 0.088mmol) and crushed 4 Å molecular sieves (20 mg) in dichloromethane (5 mL)was added formalin (0.1 mL) and the resulting suspension was stirred for30 min at room temperature. This mixture was then treated with sodiumtriacetoxyborohydride (93 mg, 0.44 mmol) and stirred an addition 15 h atroom temperature. The reaction was quenched with saturated sodiumbicarbonate solution (10 mL) and diluted with an additional 10 mL ofdichloromethane. The organic layer was separated and the aqueous layerwas washed with dichloromethane (2×20 mL). The organics were combined,dried over anhydrous sodium sulfate, filtered and evaporated underreduced pressure. The residue was purified by preparative TLC (eluant:0.5% NH₄OH/5% methanol/94.5% CH₂Cl₂) to yield 34 mg (83%) of the finalproduct. This reaction was performed the same way for the other isomers.

EXAMPLE 19

This product was prepared in an analogous fashion to that of Example 17,except Intermediate 1 was replaced with Intermediate 2. Purification bypreparative TLC (eluant: 0.5% NH₄OH/5% methanol/94.5% CH₂Cl₂) afforded203 mg (92%) as a mixture of four diastereomers. The single isomers wereobtained by purification on an HPLC equipped with a PreparativeChiralCel OD column eluting with 5% ethanol/95% hexanes with a flow rateof 9 mL/min. LC-MS for C₂₅H₃₆F₃N₃O₂ calculated 467.28, found [M+H]⁺468.3 for all 4 isomer.

EXAMPLE 20

This product was prepared in an analogous fashion to Example 17, exceptIntermediate 1 was replaced with Intermediate 5. Purification byafforded 312 mg (88%) as a mixture of four diastereomers. LC-MS forC₃₀H₃₆ClF₃N₃O₄ calculated 593.23, found [M+H]⁺ 594.3.

EXAMPLE 21

To the solution of the product described in Example 20 (286 mg, 0.482mmol) in methanol (5 mL) was added a solution of 0.5 M sodium methoxidein methanol (1.2 mL, 0.58 mmol) and the resulting mixture was stirred atroom temperature for 2 h. After completion of reaction, the mixture wasevaporated in vacuo and purified by preparative TLC (eluant: 1.0%NH₄OH/10% methanol/89% CH₂Cl₂) to yield Example 21 (201 mg, 91.6%) as amixture of four diastereomers. LC-MS for C₂₃H₃₃F₃N₃O₃ calculated 455.24,found [M+H]⁺ 456.25.

EXAMPLE 22

This product was prepared in an analogous fashion to Example 2. Thecrude product was purified by Preparative TLC (eluant: 1.0% NH₄OH/10%methanol/89% CH₂Cl₂) to afford Example 22. All four isomers wereseparately reacted to give four single compounds. LC-MS for eachdiastereomer: C₂₄H₃₅F₃N₃O₃ calculated 469.24, found [M+H]⁺ 470.3.

EXAMPLE 23

A solution of Intermediate 19 (500 mg, 1.17 mmol), Intermediate 3 (458mg, 3.51 mmol), diisopropylethylamine (407 μL, 2.34 mmol) and crushedmolecular sieves (4 Å, 250 mg) in dichloromethane (25 mL) was treatedwith sodium triacetoxyborohydride (1.24 g, 5.85 mmol) and stirred atroom temperature overnight. The reaction was quenched with saturatedsodium bicarbonate solution (25 mL) and diluted with an additional 25 mLof dichloromethane. The organic layer was separated and the aqueouslayer was washed with dichloromethane (2×20 mL). The organics werecombined, dried over anhydrous sodium sulfate, filtered and evaporatedunder reduced pressure. The residue was purified by preparative TLC(eluant: 1.0% NH₄OH/10% methanol/89% CH₂Cl₂) to yield 210 mg (86%) ofthe final product as a mixture of four diastereomers. The single isomerswere obtained by using an HPLC equipped with a Preparative ChiralCel ODcolumn eluting with 20% ethanol and 80% hexanes with a flow rate of 9mL/min. LC-MS calculated for C₂₄H₃₄F₃N₃O₃ is 469.21, found [M+H]⁺ 470.2for all 4 isomer. 3^(rd) isomer off OD ChiralCel Column: ¹H NMR (500MHz, CDCl₃) d 8.72 (s, 1H), 7.69 (s, 1H), 4.87 (br d, J=17.2 Hz, 1H),4.75 (d, J=17.4 Hz, 1H), 4.12 (dd, J=3.1, 12.4 Hz, 1H), 3.99-3.86 (m,3H), 3.47-3.39 (m, 1H), 3.41 (s, overlapped, 3H), 3.35-3.30 (m, 2H),3.20-3.08 (m, 3H), 2.87-2.80 (m, 1H), 2.62-2.54 (m, 1H), 2.16-2.02 (m,2H), 1.95 (br s, 1H), 1.88-1.81 (m, 1H), 1.78-1.57 (m, 6H), 1.41-1.32(m, 1H), 0.96 (d, J=6.7 Hz, 3H), 0.84 (d, J=6.6 Hz, 3H). 4^(th) isomeroff OD ChiralCel Column: ¹H NMR (500 MHz, CDCl₃) d¹H NMR (500 MHz,CDCl₃) d8.72 (s, 1H), 7.69 (s, 1H), 4.87 (br d, J=17.6 Hz, 1H), 4.75 (d,J=17.5 Hz, 1H), 4.10 (dd, J=3.1, 12.3 Hz, 1H), 3.99-3.88 (m, 3H),3.46-3.39 (m, 1H), 3.41 (s, overlapped, 3H), 3.35-3.30 (m, 2H),3.17-3.09 (m, 3H), 2.86-2.80 (m, 1H), 2.64-2.55 (m, 1H), 2.16-2.10 (m,1H), 2.05 (br s, 1H), 1.95-1.82 (m, 2H), 1.76-1.55 (m, 6H), 1.33-1.24(m, 1H), 0.95 (d, J=6.7 Hz, 3H), 0.83 (d, J=6.6 Hz, 3H).

EXAMPLE 24

To a solution of a single isomer described in Example 23 (100 mg, 0.203mmol) and crushed 4 Å molecular sieves (200 mg) in dichloromethane (7mL) was added formalin (0.2 mL) and the resulting suspension was stirredfor 30 min at room temperature. This mixture was then treated withsodium triacetoxyborohydride (215 mg, 1.01 mmol) and stirred an addition15 h at room temperature. The reaction was quenched with saturatedsodium bicarbonate solution (10 mL) and diluted with an additional 10 mLof dichloromethane. The organic layer was separated and the aqueouslayer was washed with dichloromethane (2×20 mL). The organics werecombined, dried over anhydrous sodium sulfate, filtered and evaporatedunder reduced pressure. The residue was purified by preparative TLC(eluant: 0.5% NH₄OH/5% methanol/94.5% CH₂Cl₂) to yield 97 mg (95%) ofthe final product. This reaction was performed the same way for theother three isomers.

EXAMPLE 25

This product was prepared in an analogous fashion to that of Example 17,except Intermediate 1 was replaced with Intermediate 4. The singleisomers were obtained by using an HPLC equipped with a PreparativeChiralCel OD column eluting with 15% ethanol and 85% hexanes with a flowrate of 9 mL/min. LC-MS for C₂₅H₃₆F₃N₃O₃ calculated 483.23, found [M+H]⁺484.2 for all four isomer.

EXAMPLE 26

This product was prepared in an analogous fashion to Example 17, exceptIntermediate 1 was replaced with Intermediate 6. LC-MS for C₂₄H₃₁F₄N₃O₂calculated 457.23, found [M+H]⁺ 458.2.

EXAMPLE 27

This product was prepared in an analogous fashion to Example 17, exceptIntermediate 1 was replaced with Intermediate 7. The single isomers wereobtained by using an HPLC equipped with a Preparative ChiralCel ODcolumn eluting with 5% ethanol and 95% hexanes with a flow rate of 9mL/min. LC-MS for C₂₄H₃₁F₆N₃O₂ calculated 507.23, found [M+H]⁺ 508.2 forall isomer.

EXAMPLE 28

A solution of Intermediate 22 (35 mg, 0.090 mmol),tetrahydro-4H-pyran-4-one (13 mg, 0.14 mmol), diisopropylethylamine (32μL, 0.18 mmol) and crushed molecular sieves (4 Å, 20 mg) indichloromethane (10 mL) was treated with sodium triacetoxyborohydride(96 mg, 0.45 mmol) and stirred at room temperature overnight. Thereaction was quenched with saturated sodium bicarbonate solution (15 mL)and diluted with an additional 15 mL of dichloromethane. The organiclayer was separated and the aqueous layer was washed withdichloromethane (2×10 mL). The organics were combined, dried overanhydrous sodium sulfate, filtered and evaporated under reducedpressure. The residue was purified by preparative TLC (eluant: 0.5%NH₄OH/5% methanol/94.5% CH₂Cl₂) to yield 30 mg (74%) of the finalproduct. LC-MS calculated for C₂₃H₃₀F₃N₃O₂ is 437.23, found [M+H]⁺438.3.

EXAMPLE 29

This product was prepared in an analogous fashion to Example 28, excepttetrahydro-4H-pyran-4-one was replaced with Intermediate 3. The singleisomers were obtained by using an HPLC equipped with a PreparativeChiralCel OD column eluting with 13% ethanol and 87% hexanes with a flowrate of 9 mL/min. LC-MS for C₂₄H₃₂F₃N₃O₃ calculated 467.23, found [M+H]⁺468.2 for all isomers.

EXAMPLE 30

A solution of intermediate 20 (641 mg, 1.60 mmol),tetrahydro-4H-pyran-4-one (220 mg, 2.24 mmol), diisopropylethylamine(279 μL, 1.60 mmol) and crushed molecular sieves (4 Å, 320 mg) indichloromethane (20 mL) was treated with sodium triacetoxyborohydride(1.70 g, 8.00 mmol) and stirred at room temperature for no longer than 5h. The reaction was quenched with saturated sodium bicarbonate solution(50 mL) and diluted with an additional 30 mL of dichloromethane. Theorganic layer was separated and the aqueous layer was washed withdichloromethane (2×30 mL). The organics were combined, dried overanhydrous sodium sulfate, filtered and evaporated under reducedpressure. The residue was purified by preparative TLC (eluant: 0.75%NH₄OH/7.5% methanol/91.75% CH₂Cl₂) to yield 626 mg (86%) of the finalproduct. ¹H NMR (500 MHz, CDCl₃) δ 8.45, (s, 3H), 7.25 (s, 1H), 4.88 (brd, J=17.4 Hz, 1H), 4.77 (d, J=17.6 Hz, 1H), 4.00-3.85 (m, 4H), 3.41 (appt, J=11.7 Hz, 2H), 3.22 (p, J=6.8 Hz, 1H), 3.13-3.07 (m, 2H), 2.82-2.74(m, 1H), 2.54-2.47 (m, 1H), 2.14 (dd, J=6.8, 12.8 Hz, 1H), 2.07-2.00 (m,1H), 1.94-1.86 (m, 2H), 1.84-1.77 (m, 3H), 1.65-1.57 (m, 2H), 1.46-1.26(m, 3H), 0.93 (d, J=6.8 Hz, 3H), 0.83 (d, J=6.8 Hz, 3H): LC-MS forC₂₃H₃₂F₃N₃O₃ calculated 455.24, found [M+H]⁺456.2.

EXAMPLE 31

To a solution of product described in Example 30 (100 mg 0.203 mmol) andcrushed 4 Å molecular sieves (150 mg) in dichloromethane (7 mL) wasadded formalin (0.2 mL) and the resulting suspension was stirred for 30min at room temperature. This mixture was then treated with sodiumtriacetoxyborohydride (215 mg, 1.01 mmol) and stirred an addition 5 h atroom temperature. The reaction was quenched with saturated sodiumbicarbonate solution (10 mL) and diluted with an additional 10 mL ofdichloromethane. The organic layer was separated and the aqueous layerwas washed with dichloromethane (2×20 mL). The organics were combined,dried over anhydrous sodium sulfate, filtered and evaporated underreduced pressure. The crude product was purified by Preparative TLC toafford Example 20 (97 mg, 95%). LC-MS for C₂₄H₃₄F₃N₃O₃ calculated469.24, found [M+H]⁺ 470.2.

EXAMPLE 32

This product was prepared in an analogous fashion to Example 30, excepttetrahydro-4H-pyran-4-one was replaced with Intermediate 1. The singleisomers were obtained by using an HPLC equipped with a PreparativeChiralCel OD column eluting with 7% ethanol and 93% hexanes with a flowrate of 9 mL/min. LC-MS for C₂₄H₃₄F₃N₃O₃ calculated 469.24, found [M+H]⁺470.2, for all four isomer.

EXAMPLE 33

This product was prepared in an analogous fashion to Example 31. Thecrude product was purified by Preparative TLC (eluant: 1.0% NH₄OH/10%methanol/89% CH₂Cl₂) to afford Example 33. All four isomers wereseparately reacted to give four single compounds. LC-MS for eachdiastereomer: C₂₅H₃₆F₃N₃O₃ calculated 483.24, found [M+H]⁺ 484.4.

EXAMPLE 34

This product was prepared in an analogous fashion to Example 30, excepttetrahydro-4H-pyran-4-one was replaced with Intermediate 2. The singleisomers were obtained by using an HPLC equipped with a PreparativeChiralCel OD column eluting with 5% ethanol and 95% hexanes with a flowrate of 9 mL/min. LC-MS for C₂₅H₃₆F₃N₃O₃ calculated 483.24, found [M+H]⁺484.2, for all four isomer.

EXAMPLE 35

This product was prepared in an analogous fashion to Example 30, excepttetrahydro-4H-pyran-4-one was replaced with Intermediate 3. The singleisomers were obtained by using an HPLC equipped with a PreparativeChiralCel OD column eluting with 21% ethanol and 79% hexanes with a flowrate of 9 mL/min. LC-MS for C₂₄H₃₄F₃N₃O₄ calculated 485.25, found [M+H]⁺486.3, for all four isomer.

EXAMPLE 36

This product was prepared in an analogous fashion to Example 31. Thecrude product was purified by Preparative TLC (eluant: 1.0% NH₄OH/10%methanol/89% CH₂Cl₂) to afford Example 36. All four isomers wereseparately reacted to give four single compounds. LC-MS for eachdiastereomer: C₂₅H₃₆F₃N₃O₄ calculated 499.24, found [M+H]⁺ 500.3.

EXAMPLE 37

Intermediate 8 (28 g, 0.10 mmol) and Intermediate 14 (25 mg, 0.088 mmol)were first dried by azeotropic distillation with toluene (3×10 mL) andplaced under high vacuum for 30 min. Under nitrogen,4-dimethylaminopyridine (7 mg, 0.053 mmol), anhydrous dichloromethane(1.0 mL), and diisopropylethylamine (30 μL, 0.175 mmol) were addedsequentially. After Intermediate 8 was in solution,bromo-tris-pyrrolidino-phosphonium hexafluorophosphate (42 mg, 0.088mmol) was added, immediately followed by additionaldiisopropylethylamine (30 μL, 0.17 mmol). The reaction mixture wasstirred at room temperature overnight and then quenched with saturatedNaHCO₃. The aqueous layer was back washed with dichloromethane (3×5 mL)and the organic layers were combined, dried over Na₂SO₄, filtered, andevaporated in vacuo. The crude product was purified by preparative TLC(eluant: 40% ethyl acetate/60% hexanes) to afford the product (21 mg,51%) as a clear film. LC-MS for C₂₄H₃₂F₃N₃O₃ calculated 467.24, found[M+H−100(Boc)]⁺ 368.2.

The product described in Step B, Example 37 (21 g, 0.045 mmol) wasdissolved in 4 N HCl in dioxane (2.0 mL) and the resulting solution wasstirred at room temperature for 1 h. The reaction was evaporated undervacuum to afford the product (20 mg, 100%) as a white powder. LC-MS forC₁₈H₂₄F₃N₃O calculated 367.20, found [M+H]⁺ 368.2.

A solution of the product described in Step B, Example 37 (20 mg, 0.045mmol), tetrahydro-4H-pyran-4-one (9 mg, 0.09 mmol),diisopropylethylamine (16 μL, 0.090 mmol) and crushed molecular sieves(4 Å, 15 mg) in dichloromethane (1.0 mL) was treated with sodiumtriacetoxyborohydride (48 mg, 0.22 mmol) and stirred at room temperatureovernight. The reaction was quenched with saturated sodium bicarbonatesolution (10 mL) and diluted with an additional 10 mL ofdichloromethane. The organic layer was separated and the aqueous layerwas washed with dichloromethane (10 mL). The organics were combined,dried over anhydrous sodium sulfate, filtered and evaporated underreduced pressure. The residue was purified by preparative TLC (eluant:0.5% NH₄OH/5% methanol/94.5% CH₂Cl₂) to yield 18 mg (88%) of the finalproduct. LC-MS for C₂₄H₃₂F₃N₃O₂ calculated 451.24, found [M+H]⁺ 452.2.

EXAMPLE 38

Intermediate 8 (250 mg, 0.81 mmol) and Intermediate 15 (280 mg, 1.00mmol) were first dried by azeotropic distillation with toluene (3×10 mL)and placed under high vacuum for 30 min. Under nitrogen,4-dimethylaminopyridine (65 mg, 0.53 mmol), anhydrous dichloromethane(3.0 mL), and diisopropylethylamine (350 μL, 2.02 mmol) were addedsequentially. After Intermediate 8 was in solution,bromo-tris-pyrrolidino-phosphonium hexafluorophosphate (378 mg, 0.810mmol) was added, immediately followed by additionaldiisopropylethylamine (350 μL, 2.02 mmol). The reaction mixture wasstirred at room temperature overnight and then quenched with saturatedNaHCO₃. The aqueous layer was back extracted with dichloromethane (3×10mL) and the organic layers were combined, dried over Na₂SO₄, filtered,and evaporated in vacuo. The crude product was purified by preparativeTLC (eluant: 20% ethyl acetate/80% hexanes) to afford the product (192mg, 50%) as a white foam. LC-MS for C₂₅H₃₄F₃N₃O₃ calculated 481.24,found [M+H−100(Boc)]⁺ 382.2.

The product described in Step B, Example 38 (100 mg, 0.21 mmol) wasdissolved with 4 N HCl in dioxane (5.0 mL) and the resulting solutionwas stirred at room temperature for 1 h. The reaction was evaporatedunder vacuum to afford the product (91 mg, 96%) as a white powder. LC-MSfor C₁₈H₂₄F₃N₃O calculated 381.20, found [M+H]⁺ 382.2.

A solution of the product described in Step B, Example 38 (91 mg, 0.20mmol), tetrahydro-4H-pyran-4-one (30 mg, 0.30 mmol),diisopropylethylamine (70 μL, 0.40 mmol) and crushed molecular sieves (4Å, 45 mg) in dichloromethane (7.0 mL) was treated with sodiumtriacetoxyborohydride (212 mg, 1.00 mmol) and stirred at roomtemperature overnight. The reaction was quenched with saturated sodiumbicarbonate solution (20 mL) and diluted with an additional 10 mL ofdichloromethane. The organic layer was separated and the aqueous layerwas washed with dichloromethane (2×10 mL). The organics were combined,dried over anhydrous sodium sulfate, filtered and evaporated underreduced pressure. The residue was purified by preparative TLC (eluant:0.5% NH₄OH/5% methanol/94.5% CH₂Cl₂) to yield 82 mg (77%) of the finalproduct. LC-MS for C₂₅H₃₄F₃N₃O₂ calculated 465.24, found [M+H]⁺ 466.2.

EXAMPLE 39

To a solution of the product described in Step A, Example 38 (100 mg,0.208 mmol) in dichloromethane (5 mL) was added 3-chloroperoxybenzoicacid (93 mg, 0.42 mmol) and the resulting solution was stirred overnightat room temperature. The mixture was cooled to 0° C. and while stirringvigorously solid calcium hydroxide was added in portions until about 1gram was added. The suspension was stirred for an additional 30 min,then filtered through celite to remove all solids. The filtrate wasevaporated in vacuo and the residue purified by preparative TLC (eluant:70% ethyl acetate/30% hexanes) to afford 79 mg (77%) of the desiredcompound. LC-MS for C₂₅H₃₄F₃N₃O₄ calculated 497.20, found [M+H]⁺ 498.2.

The product described in Step B, Example 39 (75 mg, 0.151 mmol) wasdissolved in 4 N HCl in dioxane (4 mL) and the resulting solution wasstirred at room temperature for 1 h. The reaction was evaporated undervacuum to afford the product (64 mg, 98%) as a white powder. LC-MS forC₁₈H₂₄F₃N₃O₂ calculated 397.20, found [M+H]⁺ 398.2.

To a solution of the product described in Step C, Example 39 (64 mg 0.15mmol), tetrahydro-4H-pyran-4-one (22 mg, 0.22 mmol),diisopropylethylamine (26 μL, 0.149 mmol) and crushed molecular sieves(4 Å, 30 mg) in dichloromethane (5 mL) was treated with sodiumtriacetoxyborohydride (158 mg, 0.745 mmol) and stirred at roomtemperature for no longer than 5 h. The reaction was quenched withsaturated sodium bicarbonate solution (10 mL) and diluted with anadditional 10 mL of dichloromethane. The organic layer was separated andthe aqueous layer was washed with dichloromethane (2×10 mL). Theorganics were combined, dried over anhydrous sodium sulfate, filteredand evaporated under reduced pressure. The residue was purified bypreparative TLC (eluant: 0.75% NH₄OH/7.5% methanol/91.75% CH₂Cl₂) toyield 52 mg (68%) of the final product. LC-MS for C₂₅H₃₄F₃N₃O₃calculated 482.24, found [M+H]⁺483.3.

EXAMPLE 40

A solution of intermediate 23 (65 mg, 0.14 mmol),tetrahydro-4H-pyran-4-one (26 mg, 0.28 mmol), diisopropylethylamine (25μL, 0.14 mmol) and crushed molecular sieves (4 Å, 35 mg) indichloromethane (5 mL) was treated with sodium triacetoxyborohydride(148 mg, 0.700 mmol) and stirred at room temperature overnight. Thereaction was quenched with saturated sodium bicarbonate solution (10 mL)and diluted with an additional 15 mL of dichloromethane. The organiclayer was separated and the aqueous layer was washed withdichloromethane (2×10 mL). The organics were combined, dried overanhydrous sodium sulfate, filtered and evaporated under reducedpressure. The crude product was purified by reverse phase HPLC to yieldthe final product (42 mg, 63%). LC-MS for C₂₂H₂₇F₆N₃O₂ calculated479.20, found [MH]⁺ 480.25.

EXAMPLE 41

To a solution of product described in Example 40 (40 mg 0.083 mmol) andcrushed 4 Å molecular sieves (20 mg) in dichloromethane (5 mL) was addedformalin (0.1 mL) and the resulting suspension was stirred for 30 min atroom temperature. This mixture was then treated with sodiumtriacetoxyborohydride (89 mg, 0.42 mmol) and stirred an addition 15 h atroom temperature. The reaction was quenched with saturated sodiumbicarbonate solution (10 mL) and diluted with an additional 10 mL ofdichloromethane. The organic layer was separated and the aqueous layerwas washed with dichloromethane (2×20 mL). The organics were combined,dried over anhydrous sodium sulfate, filtered and evaporated underreduced pressure. The crude product was purified by reverse phase HPLCto yield Example 41 (37.5 mg, 92%). LC-MS for C₂₃H₂₉F₆N₃O₂ [M⁺H⁺]calculated 493.22, found [MH]⁺ 494.2.

EXAMPLE 42

This product was prepared in an analogous fashion to Example 40, excepttetrahydro-4H-pyran-4-one was replaced with Intermediate 1. The singleisomers were obtained by using an HPLC equipped with a PreparativeChiralCel OD column eluting with 4% ethanol and 96% hexanes with a flowrate of 9 mL/min. LC-MS for C₂₃H₂₉F₆N₃O₂ calculated 493.22, found [M+H]⁺494.2, for all four isomer.

TABLE 1 The table below shows examples synthesized in a similar fashionto Example 40 and 41 above, where the tetrahydropyran is replaced withsome substituted tetrahydropyrans.

L-222701, L-222702, L-222703, L-222704, L-234971, L-234972, L-234973,L-234974, L-251451, L-251452 FW: formula/found Example R1 R2 Column andeluant [M + H]⁺ 43 CH₃ CH₃ Single isomers obtained from C₂₄H₃₁F₆N₃O₂Example 31 508.2 44 OMe H Preparative ChiralCel OD C₂₃H₂₉F₆N₃O₃ 93%Hexane: 7% Ethanol 510.2 45 OMe CH₃ Single isomers obtained fromC₂₄H₃₁F₆N₃O₃ Example 34 524.2 46 F H Preparative ChiralCel ODC₂₂H₂₆F₇N₃O₂ 90% Hexane: 10% Ethanol 498.1 47 CF3 H PreparativeChiralCel OD C₂₃H₂₆F₉N₃O₂ 97% Hexane: 3% Ethanol 548.3

EXAMPLE 48

A solution of intermediate 24 (250 mg, 0.558 mmol),tetrahydro-4H-pyran-4-one (90 mg, 0.84 mmol), diisopropylethylamine (100μL, 0.558 mmol) and crushed molecular sieves (4 Å, 150 mg) indichloromethane (10 mL) was treated with sodium triacetoxyborohydride(590 mg, 2.79 mmol) and stirred at room temperature for no longer than 5h. The reaction was quenched with saturated sodium bicarbonate solution(20 mL) and diluted with an additional 20 mL of dichloromethane. Theorganic layer was separated and the aqueous layer was washed withdichloromethane (2×20 mL). The organics were combined, dried overanhydrous sodium sulfate, filtered and evaporated under reducedpressure. The residue was purified by preparative TLC (eluant: 1.0%N₄OH/10% methanol/89% CH₂Cl₂) to yield 191 mg (70%) of the finalproduct. LC-MS for C₂₂H₂₇F₆N₃O₃ calculated 495.24, found [M+H]⁺ 496.2.

EXAMPLE 49

To a solution of the product described in Example 48 (150 mg 0.302 mmol)and crushed 4 Å molecular sieves (200 mg) in dichloromethane (7 mL) wasadded formalin (0.3 mL) and the resulting suspension was stirred for 30min at room temperature. This mixture was then treated with sodiumtriacetoxyborohydride (321 mg, 1.51 mmol) and stirred an addition 5 h atroom temperature. The reaction was quenched with saturated sodiumbicarbonate solution (20 mL) and diluted with an additional 20 mL ofdichloromethane. The organic layer was separated and the aqueous layerwas washed with dichloromethane (2×20 mL). The organics were combined,dried over anhydrous sodium sulfate, filtered and evaporated underreduced pressure. The crude product was purified by Preparative TLC toafford Example 49 (112 mg, 73%). LC-MS for C₂₃H₂₉F₆N₃O₃ calculated509.24, found [M+H]⁺ 510.2.

EXAMPLE 50

This product was prepared in an analogous fashion to Example 48, excepttetrahydro-4H-pyran-4-one was replaced with Intermediate 1. The singleisomers were obtained by using an HPLC equipped with a PreparativeChiralCel OD column eluting with 6% ethanol and 94% hexanes with a flowrate of 9 mL/min. LC-MS for C₂₃H₂₉F₆N₃O₃ calculated 509.24, found [M+H]⁺510.2., for all four isomer.

EXAMPLE 51

This product was prepared in an analogous fashion to Example 49 fromExample 50. The crude product was purified by Preparative TLC (eluant:1.0% NH₄OH: 10% methanol: 89% CH₂Cl₂) to afford Example 51. All fourisomers were separately reacted to give four single compounds. LC-MS foreach diastereomer: C₂₄H₃₁F₆N₃O₃ calculated 515.24, found [M+H]⁺ 516.3.

EXAMPLE 52

A solution of intermediate 25 (50 mg, 0.1 mmol),tetrahydro-4H-pyran-4-one (21 mg, 0.21 mmol), diisopropylethylamine (37μL, 0.21 mmol) and crushed molecular sieves (4 Å, 35 mg) indichloromethane (5 mL) was treated with sodium triacetoxyborohydride(111 mg, 0.525 mmol) and stirred at room temperature overnight. Thereaction was quenched with saturated sodium bicarbonate solution (10 mL)and diluted with an additional 15 mL of dichloromethane. The organiclayer was separated and the aqueous layer was washed withdichloromethane (2×10 mL). The organics were combined, dried overanhydrous sodium sulfate, filtered and evaporated under reducedpressure. The crude product was purified by preparative TLC (eluant:0.75% NH₄OH/7.5% methanol/91.75% CH₂Cl₂) to yield the final product (39mg, 75%). LC-MS for C₂₃H₂₉F₆N₃O₂ calculated 493.20, found [MH]⁺ 494.3.

EXAMPLE 53

This product was prepared in an analogous fashion to Example 52, excepttetrahydro-4H-pyran-4-one was replaced with Intermediate 3. The singleisomers were obtained by using an HPLC equipped with a PreparativeChiralCel OD column eluting with 7% ethanol and 93% hexanes with a flowrate of 9 mL/min. LC-MS for C₂₄H₃₁F₆N₃O₃ calculated 523.22, found [M+H]⁺524.4, for all four isomer.

EXAMPLE 54

This intermediate was prepared in an analogous fashion to the productdescribed in Step B, Intermediate 12, except2-iodo-1,1,1-trifluoroethane was replaced withethyl-1,1,1-trifluoroacetate. Purification by MPLC (gradient eluent0-40% ethyl acetate/hexanes) afforded the desired compound (4.26 g, 68%)as a 3:2 mixture of diastereoisomers. LC-MS for C₁₄H₂₂F₃NO₆ calculated357.70, found [M+H−100(Boc)]⁺258.1

A solution of the product described in Step A, Example 54 (4.0 g, 12mmol) in methanol (10 mL) was treated with sodium borohydride (1.34 g)and the resulting mixture was stirred at room temperature for 2 h. Thereaction was quenched with water (20 mL) and concentrated in vacuo toremove the methanol. The aqueous layer was extracted with ethyl acetate(3×50 mL) and the organics were combined, dried over sodium sulfate,filtered, and evaporated under reduced pressure. Purification by MPLC(gradient eluent 0-75% ethyl acetate/hexanes) gave the product as aclear oil. LC-MS for C₁₃₄H₂₂F₃NO₄ calculated 313.14, found[M+H−100(Boc)]⁺ 214.1

A solution of the product described in Step B, Example 54 (3.5 g, 11mmol) in chloroform/acetonitrile/water (1:1:1 solution, 63 mL) wastreated with sodium periodate (9.56 g, 44.7 mmol) and RuCl₃ trihydrate(175 mg, 0.670 mmol) and the resulting dark brown solution was stirredat room temperature for 3 h. The dark brown solution changed to a brightorange after stirring for 3 h. The mixture was diluted withdichloromethane (100 mL) and the layers were separated. The aqueouslayer was washed with dichloromethane (2×50 mL) and the organics werecombined, dried over sodium sulfate, filtered through celite, andevaporated in vacuo. Purification by flash column (gradient eluent 0-20%methanol/ethyl acetate) afforded two separated isomers as a mixture of 2diastereoisomers at the hydroxyethyl carbon. The faster eluting isomerwas the desired cis isomer (1.45 g, 40%) and the lower eluting isomerwas the undesired trans isomer (986 mg, 27%).

The faster eluting cis isomer described in Step C, Example 54 (326 mg,1.00 mmol) and Intermediate 8 (412 mg, 1.50 mmol) were first dried byazeotropic distillation with toluene (2×10 mL) and placed under highvacuum for 30 min. Under nitrogen, 4-dimethylaminopyridine (73 mg, 0.60mmol), anhydrous dichloromethane (4 mL), and diisopropylethylamine (435μL, 2.50 mmol) were added sequentially. After Intermediate 8 was insolution, bromo-tris-pyrrolidino-phosphonium hexafluorophosphate (466mg, 1.00 mmol) was added, immediately followed by additionaldiisopropylethylamine (435 μL, 2.50 mmol). The reaction mixture wasstirred at room temperature overnight and then quenched with saturatedNaHCO₃ solution. The aqueous layer was back extracted withdichloromethane (3×50 mL) and the organic layers were combined, driedover Na₂SO₄, filtered, and evaporated in vacuo. The crude product waspurified by preparative TLC (eluent: 50% ethyl acetate/50% hexanes) toafford two single isomers: higher eluting (126 mg, 25%) and lowereluting (65 mg, 13%) as yellow films. The stereochemistry of the twoisomers at the hydroxy-trifluoroethyl carbon is unknown and was notdetermined. ¹H NMR (500 MHz, CDCl₃), First higher eluting cis isomer δ8.70, (s, 3H), 7.71 (s, 1H), 5.55 (br s, 1H), 5.10 (br s, 1H), 5.0-4.86(m, 1H), 4.78 (d, J=17.6 Hz, 1H), 4.34 (br s, 1H), 4.12-4.04 (m, 1H),4.02-3.84 (m, 1H), 3.56-3.44 (m, 1H), 3.20-3.06 (m, 2H), 2.64-2.46 (m,1H), 2.44-2.29 (m, 1H), 2.26-2.16 (m, 1H), 2.12-2.00 (m, 2H), 2.00-1.82(m, 3H), 1.78-1.66 (m, 1H), 1.45 (s, 9H). ¹H NMR (500 MHz, CDCl₃),Second lower eluting cis isomer δ 8.70, (s, 3H), 7.71 (s, 1H), 5.60 (brs, 1H), 4.87 (br d, J=17.2 Hz, 1H), 4.81 (d, J=17.7 Hz, 1H), 4.29 (br s,1H), 4.20-3.93 (m, 2H), 3.20-3.05 (m, 2H), 2.72-2.56 (m, 1H), 2.26-2.18(m, 1H), 2.16-2.00 (m, 1H), 2.10-2.04 (m, 1H), 1.98-1.92 (m, 1H),1.46-1.42 (m, 1H), 1.40 (s, 9H).

The products described in step D, Example 54 (124 mg, 0.242 mmol, highereluting isomer and 60 mg, 0.117 mmol, lower eluting isomer) were eachdissolved with 4 N HCl in dioxane (5 mL) and the resulting solutionswere stirred at room temperature for 1 h. The reactions were evaporatedunder vacuum to afford the products (higher eluting isomer, 116 mg, 99%,and lower eluting isomer, 53 mg, 94%) as pale white solids. LC-MS forC₁₇H₁₉F₆N₃O₂ calculated 412.14, found [M+H]⁺ 413.15 for both isomers.

A solution of the higher eluting isomer described in Step E, Example 54(116 mg, 0.239 mmol), tetrahydro-4H-pyran-4-one (72 mg, 0.72 mmol),diisopropylethylamine (84 μL, 0.47 mmol) and crushed molecular sieves (4Å, 55 mg) in dichloromethane (5 mL) was treated with sodiumtriacetoxyborohydride (254 mg, 1.20 mmol) and stirred at roomtemperature overnight. The reaction was quenched with saturated sodiumbicarbonate solution (20 mL) and diluted with an additional 20 mL ofdichloromethane. The organic layer was separated and the aqueous layerwas washed with dichloromethane (2×10 mL). The organics were combined,dried over anhydrous sodium sulfate, filtered and evaporated underreduced pressure. The crude product was purified by preparative TLC(eluant: 1.0% NH₄OH/10% methanol/89% CH₂Cl₂) to yield the productlabeled as the higher isomer (58 mg, 49%). ¹H NMR (500 MHz, CDCl₃), δ8.73 (s, 1H), 7.70 (s, 1H), 4.86 (s, 2H), 4.17-4.11 (m, 1H), 4.06-3.95(m, 3H), 3.88 (ddd, J=7.3, 5.0, 13.0 Hz, 1H), 3.42-3.36 (m, 3H),3.21-3.07 (m, 2H), 2.77-2.70 (m, 1H), 2.60-2.52 (m, 2H), 2.29-2.21 (m,1H), 2.05 (ddd, J=6.4, 7.0, 12.0 Hz, 1H), 1.92 (dd, J=8.5, 13.0 Hz, 1H),1.86-1.78 (m, 2H), 1.44-1.30 (m, 4H). LC-MS for C₂₂H₂₇F₆N₃O₃ calculated495.20, found [M+H]⁺ 496.15.

This product was prepared in an analogous fashion to the compounddescribed in Step F, Example 54. The crude product was purified byPreparative TLC (eluant: 1.0% NH₄OH/10% methanol/89% CH₂Cl₂) to affordthe product labeled as the lower isomer (32 mg, 67%). ¹H NMR (500 MHz,CDCl₃), δ 8.71 (s, 1H), 7.69 (s, 1H), 4.87 (d, J=17.1 Hz, 1H), 4.78 (brd, J=18.0 Hz, 1H), 4.10 (dd, J=7.0, 14.2 Hz, 1H), 4.06-4.01 (m, 1H),3.97 (br d, J=11.2 Hz, 2H), 3.88 (ddd, J=5.7, 7.0, 12.7 Hz, 1H),3.46-3.35 (m, 3H), 3.22-3.07 (m, 2H), 2.76-2.66 (m, 1H), 2.62 (dd,J=6.5, 13.6 Hz, 1H), 2.55-2.48 (m, 1H), 2.14-2.00 (m, 3H), 1.84-1.76 (m,2H), 1.55 (dd, J=6.5, 6.7, 12.8 Hz, 2H), 1.35-1.26 (m, 4H). LC-MS forC₂₂H₂₇F₆N₃O₃ calculated 495.20, found [M+H]⁺ 496.15.

EXAMPLE 55

A solution of Intermediate 16 (89 mg, 0.17 mmol),tetrahydro-4H-pyran-4-one (52 mg, 0.52 mmol), diisopropylethylamine (30μL, 0.17 mmol) and crushed molecular sieves (4 Å, 200 mg) indichloromethane (6 mL) was treated with sodium triacetoxyborohydride(184 mg, 0.870 mmol) and stirred at room temperature overnight. Thereaction was quenched with saturated sodium bicarbonate solution (20 mL)and diluted with an additional 20 mL of dichloromethane. The organiclayer was separated and the aqueous layer was washed withdichloromethane (10 mL). The organics were combined, dried overanhydrous sodium sulfate, filtered and evaporated under reducedpressure. The single isomers of the crude product (65.7 mg, 78%) wereseparated by preparative TLC (CH₂Cl₂/methanol/NH₄OH/90:9:1) to yield thefinal products: Higher eluting isomer: LC-MS for C₂₂H₃₀F₃N₃O₃ calculated441.22, found [M+H]⁺ 442.30. ¹H NMR (500 MHz, CDCl₃): 8.71 (s, 1H), 7.69(s, 1H), 4.94 (d, J=17.4 Hz, 1H), 4.78 (d, J=17.40, 1H), 4.0 (m, 4H),3.40 (m, 3H), 3.12 (m, 2H), 2.80 (bs, 1H), 2.54 (m, 1H), 2.40 (m, 1H),2.00 (m, 2H), 1.85 (m, 3H), 1.14 (d, J=6.17 Hz, 3H). Lower elutingisomer: MS for C₂₂H₃₀F₃N₃O₃ calculated 441.22, found [M+H]⁺ 442.30. ¹HNMR (500 MHz, CDCl₃): 8.71 (s, 1H), 7.69 (s, 1H), 4.96 (m, 1H), 4.78 (d,J=17.40 Hz, 1H), 4.02 (m, 3H), 3.90 (m, 1H), 3.40 (t, J=11.67 Hz, 2H),3.12 (t, J=5.49 Hz, 2H), 2.84 (bs, 1H), 2.50 (ddd, J=13.04, 8.01, 4.80Hz, 1H), 2.36 (dd, J=13.50, 6.64 Hz, 1H), 2.05 (bd, J˜10 Hz, 2H), 1.88(m, 3H), 1.50 (bs, 3H), 1.16 (d, 6.41 Hz, 3H).

EXAMPLE 56

This compound was synthesized from the lower eluting isomer describedunder Example 55 using a procedure analogous to that detailed in Example2. MS for C₂₃H₃₂F₃N₃O₃ calculated 455.24, found [M+H]⁺ 456.25.

EXAMPLE 57

A solution of the amide, the synthesis of which was described underSteps A-E of Intermediate 16 (172 mg, 0.376 mmol) and3-chloroperoxybenzoic acid (191 mg, 68%, 0.752 mmol) in dichloromethane(5 mL) was stirred at room temperature for 2 h. The reaction wasquenched by the careful addition of calcium hydroxide (170 mg, 2.3mmol), and the stirring was continued for another 30 min. The solid wasfiltered off and the filtrate was evaporated to dryness to leave 156.6mg (88%) of the product, which was used in the next reaction step asobtained. MS for C₂₂H₃₀F₃N₃O₅ calculated 473.21, found [M+H]⁺ 374.30(loss of the BOC group).

The final compound was synthesized starting from the previouslydescribed N-oxide in a series of steps described in Intermediate 16,Step F, followed by the procedure detailed under Example 47, except thatthe reductive amination step was conducted at room temperature for notlonger than 2.5 h. The pure single diastereomers were obtained byseparation on chiral HPLC (ChiralCel OD, 15% ethyl alcohol in hexanes,9.0 mL/min). LC-MS for C₂₂H₃₀F₃N₃O₄ calculated 457.22, found [M+H]⁺458.20.

EXAMPLE 58

This compound was synthesized from the lower eluting diastereoisomerdescribed in Step A, Intermediate 17 according to the proceduredescribed in Example 55. LC-MS for C₂₃H₃₂F₃N₃O₃ calculated 455.24, found[M+H]⁺ 456.25.

EXAMPLE (59-62)

A solution of Intermediate 18 (first eluting isomer, 125 mg, 0.435mmol), Intermediate 8 (176 g, 0.870 mmol), 1-hydroxy-7-azobenzotriazole(60 mg, 0.435 mmol), and diisopropylethylamine (303 μL, 1.74 mmol) indichloromethane (5 mL) was treated with1-(-3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (250 mg,1.31 mmol) and the resulting mixture was stirred at room temperatureovernight. The reaction was quenched with water, and the product wasextracted into dichloromethane. The combined organic extracts were dried(anhydrous magnesium sulfate) and the solvent was removed in vacuo. Theresidue (115 mg) was separated by preparative TLC (eluant: 80% ethylacetate/20% hexanes) to yield the single isomer (the hydroxypropylside-chain, isomer 1, 65 mg, 32%) of unknown absolute stereochemistry.All four isomers were prepared as described above to give four singlecompounds labeled isomers 1-4. LC-MS for C₂₂H₃₂F₃N₃O₄ calculated 471.23,found [M+H−100(Boc)]⁺ 372.25 for all four isomers.

The product described in Step B, Example (59-62) (isomer 1, 65 mg, 0.130mmol) was dissolved in 4 N HCl in dioxane (2 mL) and the resultingsolution was stirred at room temperature for 1 h. The reaction wasevaporated under vacuum to afford the product (isomer 1, 61 mg, 99%) asa white solid. The other isomers were also prepared as described above.LC-MS for C₁₈H₂₄F₃N₃O₂ calculated 371.23, found [M+H]⁺ 372.25.

A solution of the product (isomer 1) described in Step B, Example(59-62) (61 mg, 0.14 mmol), tetrahydro-4H-pyran-4-one (27.7 mg, 0.276mmol), diisopropylethylamine (48 μL, 0.28 mmol) and crushed molecularsieves (4 Å, 50 mg) in dichloromethane (5 mL) was treated with sodiumtriacetoxyborohydride (147 mg, 0.690 mmol) and stirred at roomtemperature overnight. The reaction was quenched with saturated sodiumbicarbonate solution (20 mL) and diluted with an additional 20 mL ofdichloromethane. The organic layer was separated and the aqueous layerwas washed with dichloromethane (10 mL). The organics were combined,dried over anhydrous sodium sulfate, filtered and evaporated underreduced pressure. The crude product was purified by preparative TLC(eluant: 1.0% NH₄OH/10% methanol/89% CH₂Cl₂) to yield the final productas a single isomer of unknown absolute stereochemistry (55 mg, 80%).LC-MS for C₂₃H₃₂F₃N₃O₃ calculated 455.27, found [M+H]⁺456.3.

TABLE 2 The other three isomers synthesized in a similar fashion toExample 59 are listed in the table below.

Molecular Calculated Found Example label Formula [M] [M + H]⁺ 60 Isomer2 C₂₃H₃₂F₃N₃O₃ 455.27 456.3 2^(nd) fastest eluting isomer 61 Isomer 3C₂₃H₃₂F₃N₃O₃ 455.27 456.3 3^(rd) fastest eluting isomer 62 Isomer 4C₂₃H₃₂F₃N₃O₃ 455.27 456.3 Last eluting isomer

EXAMPLE 63

A solution of the alcohol from Intermediate 16, Step E (80 mg, 0.18mmol), triethylamine (122 μL, 0.875 mmol) and a catalytic amount of4-dimethylaminopyridine in dichloromethane (5 mL) was treated at 0° C.with neat methanesulfonyl chloride (20 μL, 0.26 mmol). The cooling bathwas removed, and the reaction mixture was stirred at room temperaturefor 1 h. It was then diluted with dichloromethane (20 mL), and washedwith water (10 mL). After drying with anhydrous sodium sulfate theorganic solvent was removed in vacuo, and the crude product (126 mg,100%) was used immediately in the next reaction step. MS forC₂₃H₃₂F₃N₃O₆S calculated 535.20, found 436.15 [M+H−BOC]⁺.

A solution of the mesylate from the previous step (126 mg, 0.175 mmol)and potassium cyanide (114 mg, 1.75 mmol) in N,N-dimethylformamide (4mL) was degassed by vacuum/nitrogen cycle and heated to 85° C.overnight. The reaction was quenched with water and the product wasextracted with a mixture of hexane/diethyl ether (8:2). The combinedextracts were dried with anhydrous magnesium sulfate and the solvent wasremoved in vacuo to yield 77.7 mg (95%) of the desired nitrile. MS forC₂₃H₂₉F₃N₄O₃ calculated 466.22, found 367.15 [M+H−BOC]⁺.

A solution of the nitrile from previous step (80 mg, 0.17 mmol) indichloromethane (10 mL) was treated with 2 mL of trifluoroacetic acidand stirred at room temperature for 2 h. The solvent was removed invacuo, and the residual trifluoroacetic acid was co-distilled withtoluene two times to yield 148 mg (100%) of the desired amine in a formof a trifluoroacetic acid salt. MS for C₁₈H₂₁F₃N₄O calculated 366.17,found 367.10 [M+H]⁺.

The final compound was synthesized according to the procedure analogousto that described under Example 55. The two respective diastereoisomerswere conveniently separated using semi-preparative HPLC on a ChiralCelOD column. The retention times under analogous analytical conditions(ChiralCel OD, 1.0 mL/min, hexanes/ethanol (85:15) were 11.23 min and18.12 min, respectively. MS for C₂₃H₂₉F₃N₄O₂ calculated: 450.22, found:451.30 [M+H]⁺.

EXAMPLE 64

This compound was synthesized in a reductive amination step analogous tothat described in Example 55. The respective diastereoisomers wereobtained by preparative TLC. MS for C₂₅H₃₄F₃N₃O₄ calculated: 497.25,found: 498.30 [M+H]⁺.

EXAMPLE 65

A solution of the product described in Step B, Intermediate 11 (5.0 g,15.2 mmol) in anhydrous tetrahydrofuran (20 mL) was added to a solutionof freshly prepared lithium diisopropylamide (19.52 mmol in 35 mL oftetrahydrofuran) at −78° C. and the resulting dark-brown mixture wasstirred for 45 min. A solution of 1-iodo-2-methylpropane (2.25 mL) wasthen added and the resulting mixture was stirred at −78° C. for 3h. Themixture was the stirred at −25° C. for 1 h (yellow solution) andquenched with saturated aqueous ammonium chloride. The mixture wasextracted with ethyl acetate (×3) and the combined organic layers werewashed with brine, dried (MgSO₄) and concentrated in vacuo. Theresulting oil was dissolved in tetrahydrofuran (30 mL) and treated with10 mL of 2 N HCl and stirred for 3 h. The aqueous tetrahydrofuran wasevaporated to afford a clear brown oil which was dissolved indichloromethane (60 mL) and treated with a saturated solution of sodiumbicarbonate (60 mL) and di-tert-butyl dicarbonate (17.7 g, 81.3 mmol).The mixture was stirred overnight and the layers were separated. Theorganic layer was washed with brine, dried (MgSO₄) and concentrated invacuo. Flash chromatography eluting with ethyl acetate/hexanes (0 to 8%)afforded 0.78 g of the cis diastereomer and 1.69 g of the trans (withsome cis) diastereomers (51%). ¹H NMR (CDCL₃, 500 MHz) □4.88-4.96(b,1H), 4.06-4.16 (b, 1H), 3.71 (s, 3H), 2.21 (m, 1H), 2.14 (d, 1H), 2.15(d, 1H), 2.06 (m, 1H), 1.85-1.92 (m, 1H), 1.72-1.79 (m, 1H), 1.58 (s,2H), 1.48-1.54 (m, 1H), 1.45 (s, 9H), 0.82-0.87 (dd, 6H).

To a solution of 0.45 g (1.5 mmol) of the cis intermediate from Step Ain tetrahydrofuran/methanol (5.0 mL) was added an aqueous solution oflithium hydroxide (0.10 g in 2.0 mL water). The mixture was stirredovernight at 60° C. and cooled to room temperature. The pH was adjustedto 7 and the methanol evaporated. The resulting suspension was extractedwith ethyl acetate (×3). The combined organic layers were washed withbrine, dried (MgSO₄) and concentrated in vacuo to afford 0.27 g (57%) ofthe title product as an oil.

The acid from Step B (0.25 g, 0.89 mmol) in dry dichloromethane (4.0 mL)at room temperature was treated with Intermediate 19 (0.49 g, 1.8 mmol)and 1-hydroxy-7-azabenzotriazole (0.12 g, 0.89 mmol). After 10 min ofstirring, 1-(−3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(0.51 g, 2.7 mmol) was added to the mixture and the reaction wasquenched with sodium bicarbonate after 18 h. The suspension wasextracted with dichloromethane (×2) and the combined organic layers weredried (MgSO₄) and concentrated in vacuo. Flash chromatograph elutingwith ethyl acetate/hexanes (15%) afforded 0.220 g (52%) of the titleproduct. LC-MS for C₂₄H₃₄F₃N₃O₃ calculated: 469.26, found 370.3 (loss ofBoc group) [M+H−100]⁺.

To a solution of the intermediate from Step C (0.220 g) in ethyl acetate(1.0 mL) was added a saturated solution of HCl in ethyl acetate and themixture was stirred for 30 min. Removal of the volatiles in vacuo gavethe desired product as the HCl salt. LC-MS for C₁₉H₂₆F₃N₃O calculated:369.20, found: 370.2 [M+H]⁺.

A solution of the intermediate from Step D (0.190 g, 0.468 mmol) indichloromethane (3.0 mL) and diisopropylethylamine (0.123 mL) wastreated with tetrahydro-4H-pyran 4-one (0.065 mL, 0.70 mmol) and 4 Åmolecular sieve. After stirring the mixture for 45 min, sodiumtriacetoxyborohydride (0.198 g, 0.936 mmol) was added. The mixture wasstirred for 18 h, filtered and the filtrate was extracted with water.The organic layer was dried (MgSO₄) and concentrated in vacuo. Reversephase HPLC purification of the crude afforded the title product whichwas subsequently transformed to the HCl salt (0.072 g). LC-MS forC₂₄H₃₄F₃N₃O₂ calculated: 453.26, found 454.25 [M+H]⁺.

EXAMPLE 66

A solution of the intermediate in Step C, Example 65 (00.10 g, 0.23mmol) in chloroform (20 mL) was treated with 3-chloroperoxybenzoic acid(0.198 g, 1.15 mmol) and the mixture was stirred for 16 h. The solventwas evaporated and flash chromatography eluting with ethylacetate/hexanes (75%) afford 0.060 g of the N-oxide title compound(54%). LC-MS for C₂₄H₃₄F₃N₃O₄ calculated: 485.25, found 386.3 (loss ofBoc group) [M+H−100]⁺.

To a solution of the intermediate from Step A in ethyl acetate (1.0 mL)was added a saturated solution of HCl/ethyl acetate. The resultingsolution was stirred for 30 min. Removal of the volatiles under vacuumgave the desired product as the HCl salt. LC-MS for C₁₉H₂₆F₃N₃O₂calculated: 385.20, found: 386.3 [M+H]⁺.

A solution of the intermediate from Step B (0.053 g, 0.13 mmol) indichloromethane (2.0 mL) and diisopropylethylamine (0.033 mL) wastreated with tetrahydro-4H-pyran 4-one (0.017 mL, 0.19 mmol) and 4 Åmolecular sieve. After stirring the mixture for 45 min, sodiumtriacetoxyborohydride (0.053 g, 0.25 mmol) was added. The mixture wasstirred for 18 h, filtered and the filtrate was extracted with water.The organic layer was dried (MgSO₄) and concentrated in vacuo. Reversephase HPLC purification of the crude afforded gave the title product,which was subsequently transformed to the HCl salt (0.029 g). LC-MS forC₂₄H₃₄F₃N₃O₃ calculated: 469.26, found: 470.2 [M+H]⁺.

EXAMPLE 67

A solution of methyl phenylacetate (15 g, 99 mmol) in tetrahydrofuran(200 mL) and N,N′-dimethylpropyleneurea (50 ml) at 0° C. was treatedwith sodium hydride (7.99 g, 199 mmol) and the mixture stirred for 2 hat 50° C. (hydrogen gas evolution). After cooling to room temperaturecis-1,4-dichloro-2-butene was added to the mixture (exothermic reaction)and the mixture was stirred at 50° C. for 3 h. The mixture was cooled toroom temperature, quenched with saturated ammonium chloride andextracted with ethyl acetate (×2). The combined organic layers werewashed with brine, dried (MgSO₄) and concentrated. Flash chromatographyeluting with 3% ethyl acetate in hexanes afforded 7 g of the titleproduct. ¹H NMR (CDCl₃, 500 MHz) d7.35 (m, 5H), 5.78 (s, 2H), 3.65 (s,3H), 3.42 (d, 2H), 2.78 (d, 2H).

A solution of the intermediate from Step A (2.0 g, 9.9 mmol) intetrahydrofuran (5 mL) at 0° C. was treated with 4.94 mL of 1.0 Mborane-tetrahydrofuran complex. The mixture was stirred at roomtemperature overnight and quenched with 5 mL of water. Borax (2.28 g,14.8 mmol) was added to the mixture and after 18 h the mixture wasdiluted with water and extracted with ethyl acetate (×2). The organiclayer was dried (MgSO₄) and concentrated. Flash chromatography elutingwith ethyl acetate/hexanes (15%) in hexanes afforded 1.2 g of acis/trans mixture of the title alcohol.

A solution of the intermediate in Step B (1.2 g, 5.4 mmol) in acetone(5.0 mL) was treated with 2 mL of Jones' reagent (10.3 g CrO₃ in 35 mLwater and 8.8 mL of H₂SO₄) and the mixture was stirred for 2 h. Theacetone was evaporated and the residue diluted with ethyl acetate andextracted with water (×3). The organic layer was washed with brine,dried (MgSO₄) and concentrated in vacuo. Column chromatography elutingwith ethyl acetate/hexanes (10-20%) afforded 0.34 g of the title ketone.¹H NMR (CDCl₃, 500 MHz) 7.35 (m, 5H), 3.68 (s, 3H), 3.27 (d, 2H), 3.0(m, 2H), 2.65 (d, 2H).

To a solution of the intermediate from Step C (0.19 g, 0.87 mmol) intetrahydrofuran/methanol (5.0 mL) was added an aqueous solution oflithium hydroxide (0.074 g in 2.0 mL water). The mixture was stirred for6 h at 60° C. and cooled to room temperature. The pH was adjusted to 7and the methanol evaporated. The resulting suspension was extracted withethyl acetate (×3). The combined organic layers were washed with brine,dried (MgSO₄) and concentrated in vacuo to afford 0.145 g (82%) of thetitle product as an oil. ¹H NMR (CDCl₃, 500 MHz) d7.38 (m, 5H), 3.28 (d,2H), 3.05 (m, 2H), 2.63 (d, 2H).

The acid from Step D (0.14 g, 0.71 mmol) in dry dichloromethane (3.0 mL)at room temperature was treated with Intermediate 8 (0.39 g, 1.4 mmol)and 1-hydroxy-7-azabenzotriazole (0.096 g, 0.71 mmol). After 10 min ofstirring, 1-(−3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(0.4 g, 2 mmol) was added to the mixture and the reaction was quenchedwith sodium bicarbonate after 18 h. The suspension was extracted withdichloromethane (×2) and the combine organic layers dried (MgSO₄) andconcentrated in vacuo. Flash chromatograph eluting with ethylacetate/hexanes (25-30%) afforded 0.212 g (77%) of the title product.LC-MS for C₂₁H₁₉F₃N₂O₂ [M+H]⁺ calculated 389.14, found 389.05.

A solution of the intermediate from Step E (0.2 g, 0.5 mmol) indichloromethane (4.0 mL) was treated with tetrahydro-2H-pyran-4-ylamine(0.105 g, 0.76 mmol) and 4 Å molecular sieve. After stirring the mixturefor 45 min, sodium triacetoxyborohydride (0.216 g, 1.02 mmol) was added.The mixture was stirred for 18 h, filtered and the filtrate wasextracted with water. The organic layer was dried (MgSO₄) andconcentrated in vacuo. Column chromatography eluting with methanol/ethylacetate (3%) afforded 0.082 g of the two cis products and 0.020 g of thetwo trans title products which were subsequently transformed to theirHCl salts. LC-MS for C₂₆H₃₀F₃N₃O₂ calculated 473.23, found 474.25[M+H]⁺.

EXAMPLE 68

To a solution of the product described in Intermediate 28, Step C (1.44g, 5.68 mmol) in 48% HBr (12 mL), was added copper(I) bromide (1.01 g,7.04 mmol), and saturated sodium nitrite (520 mg, 7.55 mmol) solution.The reaction mixture was stirred at room temperature for 1 h and heatedat 100° C. for 20 min. The reaction mixture was made alkaline by theaddition of 50% KOH solution, and then extracted with ethyl acetate(four times). The organic portions were combined, washed by water andbrine, dried over Na₂SO₄, filtered and concentrated. The residue waspurified by column chromatography (silica gel, 50% ethyl acetate/hexanesto 75% ethyl acetate/hexanes) to yield the title compound (Example 52,Step A, 0.79 g, 52%). ESI-MS calculated For C₁₅H₁₃BrN₂O: 316.02; Found:[M+H] 317.

A solution of the amide intermediate from Example 68, Step A (0.79 g) inconcentrated HCl (50 mL) was refluxed for 24 h. The solvent and extraHCl were evaporated under vacuum. The residue was then suspended withCa(OH)₂ (1.20 g) in dichloromethane (100 mL) and stirred at roomtemperature for 12 h. The solid precipitate was filtered and thefiltrate was concentrated to give the title compound (Example 55, StepB, 274 mg, 53%). ¹H-NMR (500 MHz, CD₃OD) δ 8.46 (s, 1H), 7.48 (s, 1H),4.00 (s, 2H), 3.23 (t, J=6.0 Hz, 2H), 2.80 (t, J=6.0 Hz, 2H).

To a flask was added Intermediate 11 (274 mg, 1.01 mmol), the productdescribed in Step B, Example 68 (215 mg, 1.01 mmol),bromo-tris-pyrrolidino-phosphonium hexafluorophosphate (471 mg, 1.01mmol), 4-dimethylaminopyridine (74 mg, 0.61 mmol), diisopropylethylamine(528 μL, 3.03 mmol) and dichloromethane (5 mL). The resulting mixturewas stirred for 36 h under nitrogen, diluted by dichloromethane, washedby water (adding 1 mL of 1N HCl) and brine, dried over Na₂SO₄, filteredand concentrated. The residue was purified by flash columnchromatography (silica gel, 50% ethyl acetate/hexanes to 80% ethylacetate/hexanes) to yield the title compound (Example 55, Step C) (358mg, 93%). ¹H-NMR (400 MHz, CDCl₃) δ 8.52 (s, 1H), 7.61 (s, 1H),4.67-4.85 (m, 3H), 3.82-4.00 (m, 3H), 3.00 (m, 2H), 1.60-2.20 (m, 7H),1.44 (s, 9H), 0.89 (m, 6H). ESI-MS calculated For C₂₂H₃₂BrN₃O₃: 465.16;Found: [M+H and M+H+2]⁺ 466 and 468 respectively.

A solution of the Boc amide intermediate from Example 68, Step C (350mg, 0.75 mmol) in 4.0 N HCl/dioxane (4.0 mL, 16 mmol) was stirred for 12h. Solvent was evaporated to yield the product as an HCl salt (342 mg,100%). ¹H-NMR (500 MHz, CD₃OD) δ 8.50 (s, 1H), 7.60 (s, 1H), 4.72-4.78(m, 2H), 3.86-3.92 (m, 2H), 3.75 (s, 3H), 3.29 (m, 1H), 3.00 (m, 2H),2.50 (m, 1H), 2.02-2.14 (m, 2H), 1.78-1.92 (m, 2H), 1.62-1.69 (m, 1H),1.28-1.35 (m, 1H), 0.88 (m, 6H). ESI-MS calculated For C₁₇H₂₄BrN₃O:365.11; Found: [M+H]⁺ 366.

A solution of the product described in Step D, Example 68 (55 mg, 0.13mmol), tetrahydro-4H-pyran-4-one (35 mg, 0.38 mmol),diisopropylethylamine (44 μL, 0.25 mmol) and crushed molecular sieves (4Å, 150 mg) in dichloromethane (4 mL) was treated with sodiumtriacetoxyborohydride (132 mg, 0.625 mmol) and stirred at roomtemperature overnight. The reaction was quenched with saturated sodiumbicarbonate solution (10 mL) and diluted with an additional 15 mL ofdichloromethane. The organic layer was separated and the aqueous layerwas washed with dichloromethane (2×10 mL). The organics were combined,dried over anhydrous sodium sulfate, filtered and evaporated underreduced pressure. The residue was purified on preparative TLC (1000micron) (eluent: 1.0% NH₄OH/10% methanol/89% CH₂Cl₂) to yield the finalproduct of the title compound as a free base. Its HCl salt (47.9 mg) wasformed by treatment with 4 N HCl/dioxane. ¹H NMR (CDCl₃, 500 MHz): d8.50 (s, 1H), 7.60 (s, 1H), 4.68-4.78 (m, 2H), 3.97 (m, 2H), 3.88 (m,2H), 3.38-3.48 (m, 2H), 3.18 (m, 1H), 2.98 (m, 2H), 2.75 (m, 1H), 2.52(m, 1H), 2.15 (m, 1H), 2.04 (br s, 1H), 1.74-1.93 (m, 4H), 1.24-1.60 (m,5H), 0.88 (m, 6H). LC-MS calculated for C₂₂H₃₂BrN₃O₂: 449.17; Found:[M+H and M+H+2]⁺ 450 and 452 respectively.

EXAMPLE 69

This product was prepared in an analogous fashion to that of Example 68,except tetrahydro-4H-pyran-4-one was replaced with Intermediate 3. ¹HNMR (CDCl₃, 500 MHz): d 8.51 (s, 1H), 7.60 (s, 1H), 4.66-4.80 (m, 2H),4.11 (m, 1H), 3.86-3.98 (m, 3H), 3.41 (s, 3H), 3.34 (m, 2H), 3.16 (m,1H), 2.99 (m, 2H), 2.84 (br s, 1H), 2.58 (m, 1H), 1.56-2.14 (m, 7H),1.34 (m, 1H), 0.88 (m, 6H). LC-MS calculated for C₂₃H₃₄BrN₃O₃: 479.18;Found: 480 and 482 (M+H and M+H+2).

EXAMPLE 70

A solution of 3,5-dinitro-1-methyl-2-pyridone (5.40 g, 27.1 mmol) andtert-butyl 4-oxo-1-piperidinecarboxylate (6.48 g, 32.5 mmol) in 2 MNH₃/methanol (100 mL) was stirred at 60° C. for 16 h. The solvent wasevaporated under vacuum and the residue was purified by flash columnchromatography (silica gel, 30% ethyl acetate/hexanes to 50% ethylacetate/hexanes) to yield the title compound (6.22 g, 82%). ¹H-NMR (500MHz, CDCl₃) δ 9.26 (d, J=2.5 Hz, 1H), 8.24 (d, J=2.5 Hz, 1H), 4.72 (s,2H), 3.81 (t, J=6.0 Hz, 2H), 3.14 (t, J=6.0 Hz, 2H), 1.52 (s, 9H). LC-MScalculated For C₁₃H₁₇N₃O₄: 279.12; Found: [M+H]⁺ 280.

A solution of the intermediate described in Example 70, Step A (6.15 g,22.02 mmol) in 4 N HCl/dioxane (110 mL, 440 mmol) was stirred for 12 h.Solvent was evaporated to yield the compound as the HCl salt (5.47 g,99%). ¹H-NMR (400 MHz, CD₃OD) δ 9.32 (d, J=2.5 Hz, 1H), 8.59 (d, J=2.5Hz, 1H), 4.57 (s, 2H), 3.71 (t, J=6.0 Hz, 2H), 3.38 (t, J=6.0 Hz, 2H).

To a solution of Intermediate 9 (8.39 g, 23.9 mmol) in dichloromethane(80 mL), was added 2 M oxalyl chloride in dichloromethane (17.36 mL,34.72 mmol) and N,N-dimethylformamide (˜100 μL). The reaction mixturewas stirred for 3 h, and concentrated. The residue was put on highvacuum for 2 h and dissolved in dichloromethane (40 mL). The formed acidchloride was added into a solution of the product described above(Example 70, Step B, 5.47 g, 21.70 mmol) and diisopropylethylamine(13.61 mL, 78.12 mmol) in dichloromethane (40 mL) at 0° C. The reactionmixture was stirred for 16 h and diluted with dichloromethane, washedwith 10% NaHCO₃ and brine, dried over Na₂SO₄, filtered and concentrated.The residue was purified by flash column chromatography (silica gel, 80%ethyl acetate/hexane to 100% ethyl acetate) to yield the title compound(7.25 g, 65.3%). LC-MS calculated For C₂₄H₃₁F₃N₄O₅: 512.22; Found:[M+H]⁺ 513.

To a solution of intermediate described in Example 70, Step C (7.22 g,14.1 mmol) in ethanol (150 mL) was added 10% Pd/C (750 mg). The reactionmixture was placed in a Parr apparatus and shaken under 50 lb pressureof H₂ for 2 h. The solution was filtered through celite and concentratedunder vacuum to yield the desired product (6.97 g, 100%). LC-MScalculated For C₂₄H₃₃F₃N₄O₃: 482.25; Found: [M+H]⁺ 483.

This compound was prepared starting from the intermediate described inExample 70, Step D as detailed in Example 52, Step A. LC-MS calculatedFor C₂₄H₃₁BrF₃N₃O₃: 545.15; Found:[M+H]⁺ 546.

To a mixture the product described in Example 70, Step E (150 mg, 0.275mmol), palladium (II) acetate (1 mg), phenylboronic acid (36.8 mg, 0.302mmol), K₂CO₃ (190 mg, 1.38 mmol) and tetrabutylammonium bromide (88.7mg, 0.275 mmol) was added slowly water (1 mL) under nitrogen. Thereaction mixture was stirred and heated at 70° C. for 10 h, diluted withwater (5 mL) and extracted with ethyl acetate (five times). The organicportions were combined, washed by brine, dried over Na₂SO₄, filtered andconcentrated. The residue was purified by preparative TLC (silica gel,1000 micron) (developed by 80% ethyl acetate/hexanes) to yield theproduct (124 mg, 83%). ¹H-NMR (500 MHz, CDCl₃) δ 8.68 (m, 1H), 7.42-7.66(m, 6H), 4.75-4.98 (m, 2H), 3.88-4.20 (m, 6H), 3.22-3.62 (m, 2H), 3.10(s, 2H), 2.74-2.86(m, 2H), 2.44 (m, 1H), 1.80-2.21 (m, 4H), 1.53-1.74(m,3H), 0.94(m, 6H). LC-MS calculated For C₃₀H₃₆F₃N₃O₃: 543.27; Found:[M+H]⁺ 544.

To a solution of the product described in Example 70, Step F (120 mg,0.221 mmol) in ethanol (10 mL) was added sodium borohydride (168 mg,4.42 mmol). The reaction mixture was stirred for 16 h, and diluted withmethanol. The extra sodium borohydride was destroyed by 4 N HCl indioxane and then the solvent was evaporated under vacuum. The residuewas purified by preparative TLC (silica gel, 1000 micron) (developed by10% [aqueous NH₄OH/methanol 1/9]/dichloromethane) to yield the finalproduct of the title compound as a free base. Its HCl salt (49.0 mg) wasformed by treatment with 4 N HCl/dioxane. ¹H NMR (400 MHz, CDCl₃): d8.67 (s, 1H), 7.40-7.62 (m, 6H), 4.83 (s, 2H), 3.94-4.02 (m, 4H),3.33-3.44 (m, 3H), 2.96-3.12 (m, 3H), 2.56 (m, 1H), 1.84-2.18 (m, 6H),1.55-1.70 (m, 4H), 1.26 (s, 1H), 0.86-0.93 (m, 6H). LC-MS calculated forC₂₈H₃₇N₃O₂: 447.29; Found: [M+H]⁺ 448.

TABLE 3 The table below shows other examples synthesized in a similarfashion to Example 70. The difference is the replacement of the phenylsubstituent with various substituted aryl groups Molecular CalculatedExample substituent Formula [M] Found [M + H]⁺ 71

C₂₉H₃₉N₃O₂: 461.30 462.3 72

C₂₈H₃₆N₃O₂F 465.27 466.3 73

C₂₉H₃₉N₃O₃ 477.30 478.3 74

C₂₉H₃₆N₃O₂F₃ 515.24 516.3 75

C₂₉H₃₆N₃O₂F₃ 515.24 516.3 76

C₂₈H₃₅N₃O₂F₂ 483.26 484.3 77

C₂₈H₃₅N₃O₂F₂ 483.26 484.3 78

C₂₈H₃₅N₃O₂F₂ 483.26 484.3 79

C₂₇H₃₆N₄O₂ 448.27 449.3 80

C₂₇N₃₆N₄O₂ 448.27 449.3 81

C₂₇H₃₆N₄O₂ 448.27 449.3 82

C₂₈H₃₈N₄O₃ 478.28 479.3

EXAMPLE 83

A solution of the lower eluting isomer described in Example 55 (40 mg,0.091 mmol), acetic acid (210 μL, 3.62 mmol) in tetrahydrofuran (6 mL)was added to a solution containing nBu₃P (900 μL, 3.62 mmol) and diethylazodicarboxylate (570 μL, 3.62 mmol) in tetrahydrofuran (6 mL) at 0° C.,and stirring was continued at room temperature for 6 h. The reactionmixture was diluted with dichloromethane, washed with sodiumbicarbonate, dried with anhydrous sodium sulfate and the solvent wasremoved in vacuo. Preparative TLC purification(dichloromethane/methanol/ammonium hydroxide: 90:9:1) gave 13.5 mg (31%)of the desired product. LC MS: for C₂₄H₃₂N₃F₃O₄ calculated 483.23, found484.30 [M+H]⁺ .

EXAMPLE 84

A mixture of 9.70 g (97.0 mmol) of tetrhydro-4H-pyran-4-one and 10.5 g(150 mmol) of pyrrolidine was stirred at room temperature for 1.5 h. Theexcess pyrrolidine was removed under vacuum. The residue was dissolvedin 90 mL of diethyl ether, cooled to 0° C. and 7.4 mL of acrolein wasadded. The resulting mixture was stirred at room temperature overnight.67 mL of water was added, followed by a solution of 14 g of sulfuricacid (98%) in 33 mL of water. The ether and 10 mL of water were removedunder reduced pressure, the remaining mixture was refluxed for 0.3 h andthen cooled to room temperature. The resulting dark mixture wasextracted with dichloromethane (4×100 mL), dried over anhydrous Na₂SO₄and evaporated. The residue was purified by MPLC (30% ethylacetate/hexanes). A mixture (6.6 g) of endo/exo isomers (˜1/1) wasobtained together with pure fast isomer (1.0 g, endo) and pure slowisomer (0.8 g, exo). ¹H NMR (400 MHz, CDCl₃): δ endo: 4.58 (d, J=11.6Hz, 1H), 4.20 (d, J=11.2 Hz, 1H), 4.17 (d, J=11.2 Hz, 1H), 3.91 (d,J=11.3 Hz, 1H), 3.72 (d, J=11.5 Hz, 1H), 2.60-2.30 (m, 4H), 2.13 (m,1H), 2.02 (m, 1H), 1.80 (m, 1H). Exo: 4.54 (d, J=1.1 Hz, 1H), 4.10 (dd,J=11.4 Hz, 2H), 3.80 (dd, J=11.5 Hz, 2H), 2.86 (s, 1H), 2.70 (m, 1H),2.50 (s, 1H), 2.38 (m, 2H), 2.10 (m, 1H), 1.78 (m, 1H).

To a mixture of the hydroxyketone from Step A, Example 84 (endo/exo:˜1:1, 3.12 g, 20 mmol), 1,8-diazabicyclo[5.4.0]undec-7-ene (9.0 g, 60mmol) in benzene (25 mL) at 0° C. was added dropwise a neat solution oftrifluoromethanesulfonic anhydride. An exothermic reaction was observed.The reaction mixture was stirred for 1 h, poured directly onto a silicagel column, eluting with 20% Et₂O/hexanes. The desired product wasobtained as a light yellow oil (1.80 g). ¹H NMR (400 MHz, CDCl₃): δ 5.98(m, 1H), 5.65 (m, 1H), 4.10 (dd, 1H), 3.90(dd, 1H), 3.78 (dd, 1H), 3.65(dd, 1H), 2.80 (m, 3H), 2.50 (d, J=11.5).

A mixture of the unsaturated ketone from Step B, Example 84 (9.0 g) and10% Pd/C (0.9 g) in 50 mL of ethyl acetate was hydrogenated on a Parrapparatus for 2 h under 50 lb of hydrogen. The catalyst was removed byfiltration. The filtrate was evaporated. The product was obtained as alight yellow solid (6.817 g). ¹H NMR (400 MHz, CDCl₃): δ 4.24 (d, J=11.5Hz, 2H), 3.90 (d, J=11.60 Hz, 2H), 2.58 (m, 1H), 2.38 (br S, 2H), 2.25(m, 2H), 2.08 (m, 2H), 1.58 (m, 1H).

Intermediate 19 (117 mg, 0.300 mmol), the bicyclic ketone from Step C,Example 84 (84 mg, 0.6 mmol), diisopropylethylamine (78 mg, 0.60 mmol),molecular sieves (4 Å, 200 mg) and sodium triacetoxyborohydride (125 mg,0.600 mmol) were mixed with 10 mL of dichloromethane. The mixture wasstirred for 5 h, LC-MS showed a complete conversion. The reaction wasquenched with saturated aqueous sodium carbonate, filtered to removeinsoluble molecular sieves. The organic phase was separated and driedover sodium sulfate. The crude product was purified on preparative TLC(10%[aqueous NH₄OH/methanol 1/9]/dichloromethane) to yield the titlecompound as a white solid (70 mg, 44%). ESI-MS calculated forC₂₆H₃₆F₃N₃O₂: 479; Found: 480 [M+H]⁺. The endo and exo single isomerswere obtained by using an HPLC equipped with a preparative ChiralCel ODcolumn eluting with 10% ethanol and 90% hexanes with a flow rate of 9mL/min.

EXAMPLE 85

This compound was prepared starting from Intermediate 20 according tothe procedure as detailed in Example 84. ESI-MS calculated forC₂₆H₃₆F₃N₃O₃: 495; Found: 496 [M+H]⁺. The endo and exo single isomerswere obtained by using an HPLC equipped with a preparative ChiralCel ODcolumn eluting with 10% ethanol and 90% hexanes with a flow rate of 9mL/min.

EXAMPLE 86

A mixture of tetrahydro-4H-pyran-4-one (10 g, 0.10 mol) and pyrrolidine(12 g, 0.15 mol) was stirred for 2 h. Excessive pyrrolidine was removedunder vacuum. The enamine residue was dissolved in 50 mL ofacetonitrile. To this solution was added a neat solution of methylα-bromomethyl acrylate. The mixture was stirred for 2 h before water (30mL) was added. After being stirred for additional 2 h, acetonitrile wasremoved under vacuum. The crude product was extracted into ethyl acetate(3×) and dried over sodium sulfate. Flash chromatography (50% ethylacetate/hexanes) afforded three components. The most polar componentcontained the desired product. Further purification on MPLC (30% ethylacetate/hexanes) afforded the pure compound (3.4 g, 17%). ¹H NMR (400MHz, CDCl₃): δ 4.20, 4.18 (ss, 2H),3.76 (s, 3H), 3.70 (s, 2H), 3.00,2.98 (ss, 2H), 2.61 (m, 1H), 2.38 (m, 2H), 2.22 (m, 2H).

This compound was prepared starting from Intermediate 19 and the ketoester from Step A, Example 86 according to the procedure detailed inExample 84. ESI-MS calculated for C₂₈H₃₈F₃N₃O₄: 537; Found: 538 [M+H]⁺.The endo and exo single isomers were obtained by using an HPLC equippedwith a preparative ChiralCel OD column eluting with 10% ethanol and 90%hexanes with a flow rate of 9 mL/min.

EXAMPLE 87

To a stirred solution of 1-morpholino-1-cyclopentene (15.3 g, 100 mmol)in ether (150 mL) at 0° C. was added acrolein neat solution (90%, 9 mL).The resulting mixture was stirred at room temperature overnight, mixedwith water (70 mL) and a solution of concentrated sulfuric acid (15 mL)in 30 mL of water. The ether was removed and the aqueous solution wasrefluxed for 30 min. The dark solution was cooled to room temperature,extracted with dichloromethane (3×), dried over sodium sulfate. Flashchromatography (50% ethyl acetate/hexanes) afforded two components. Fastisomer (3.2 g): ¹H NMR (400 MHz, CDCl₃): δ 4.05 (m, 1H), 2.44 (m, 1H),2.24 (m, 1H), 2.10-1.60 (m, 9H). Slow isomer (3.0 g): ¹H NMR (400 MHz,CDCl₃): δ 4.35 (m, 1H), 2.40-1.60 (m, 11H).

This compound as a mixture of all diastereoisomers was prepared startingfrom Intermediate 19 and the hydroxy ketone (fast or slow isomers, StepA, Example 87) according to the same procedure as detailed in Example71. ESI-MS calculated for C₂₅H₃₆F₃N₃O₂: 479; Found: 480 [M+H]⁺. Thecorresponding major single isomers from the fast and slow eluted hydroxyketones were obtained by using an HPLC equipped with a preparativeChiralCel OD column eluting with 5% ethanol and 95% hexanes with a flowrate of 9 mL/min.

EXAMPLE 88

To a stirred solution of 3,4-anhydroerythritol (25 g, 240 mmol) intetrahydrofuran (100 mL) was added lithium hydride (2.1 g, 260 mmol) at0° C. The resulting suspension was stirred for 1 h before iodomethane(15.6 mL, 250 mmol) was added. The suspension was stirred for 2 days,then at 50° C. for 2 h. After being cooled to room temperature, thereaction was quenched with ice water, extracted with ethyl acetate (3×),dried over sodium sulfate and evaporated. The residue was purified byflash chromatogrpahy (10% methanol/dichloromethane) to yield the desiredmono alcohol contaminated with over alkylated ether. Further flashchromatography (eluant: ethyl acetate) afforded the pure alcohol as acolorless oil (3.0 g, 11%). ¹H NMR (400 MHz, CDCl₃): δ 4.29 (m, 1H),3.90 (m, 3H), 3.78 (m, 2H), 3.42 (s, 3H), 2.60 (br s, 1H).

To a stirred solution of 2 M oxalyl chloride (12 mL, 24 mmol) was addeddichloromethane (10 mL). The solution was cooled at −78° C. undernitrogen, dimethyl sulfoxide was added (2.83 mL, 40.0 mmol) dropwise,stirred for 10 min, then solution of the alcohol from Step A, Example 88(2.36 g, 20 mmol) was added in dichloromethane (10 mL). The resultingmixture was stirred for 30 min before triethylamine (11.5 mL, 80 mmol)was added. After being warmed to room temperature, the mixture wasdiluted with ether. The resulting solid was removed by filtration andthe filtrate was concentrated and the residue purified by flashchromatography (eluant: 1:1 ether/dichloromethane) to yield the desiredketone as a yellow oil (2.4 g, 100%). ¹H NMR (400 MHz, CDCl₃): δ 4.40(m, 1H), 4.00 (m, 1H), 3.90 (m, 1H), 3.58 (s, 3H).

This compound was prepared starting from Intermediate 19 and thetetrahydrofuran ketone from Step B, Example 88 according to theprocedure detailed in Example 71. LC-MS calculated for C₂₃H₃₂F₃N₃O₃:455; Found: 456 [M+H]⁺. The two major cis single isomers were obtainedby using an HPLC equipped with a preparative ChiralCel OD column elutingwith 10% ethanol and 90% hexanes with a flow rate of 9 mL/min.

EXAMPLE 89

This compound was prepared starting from Intermediate 19 andcommercially available 2-methyl tetrahydrofuran-3-one according to theprocedure detailed in Example 84. LC-MS calculated for C₂₃H₃₂F₃N₃O₂:439; Found: 440 [M+H]⁺. The two major cis single isomers were obtainedby using an HPLC equipped with a preparative ChiralCel OD column elutingwith 5% ethanol and 95% hexanes with a flow rate of 9 mL/min.

EXAMPLE 90

Intermediate 19 (76 mg, 0.18 mmol) was combined with Intermediate 26 (18mg, 0.14 mmol), N,N-diisopropylethylamine (74 μL, 0.43 mmol), 4 Åpowdered molecular sieves (100 mg), and sodium triacetoxyborohydride(150 mg, 0.710 mmol) in 5 mL of dichloromethane. The reaction mixturewas stirred at room temperature for 3 days, then filtered throughcelite, washed with saturated sodium bicarbonate and brine, and driedover Na₂SO₄, filtered and concentrated under reduced pressure. Theproduct was purified first by preparative TLC (silica gel, 0.5%NH₄OH/4.5% methanol/95% dichloromethane) to give a crude product ofwhich 20% was purified by reverse phase HPLC (C18, 20-100% MeCN/H₂O) andconverted to its hydrochloride salt by the addition of hydrogen chloride(2 M solution in ethyl ether) to give 2.2 mg of a white solid (17%).ESI-MS calculated for C₂₅H₃₄F₃N₃O₂: 465.26; found 466 [M+H]⁺.

EXAMPLE 91

Intermediate 20 (129 mg, 0.32 mmol) was combined with Intermediate 26(40 mg, 0.32 mmol), N,N-diisopropylethylamine (175 μL, 1.05 mmol), 4 Åpowdered molecular sieves (100 mg), and sodium triacetoxyborohydride(268 mg, 1.27 mmol) in 5 mL dichloromethane. The reaction mixture wasstirred at room temperature for 1.5 h and then was placed in therefrigerator overnight before being filtered through celite, washed withsaturated sodium bicarbonate and brine, and dried over Na₂SO₄, filteredand concentrated under reduced pressure. The product was purified bypreparative TLC (silica gel, 0.5% NH₄OH/4.5% methanol/95%dichloromethane) and converted to its hydrochloride salt by the additionof hydrogen chloride (2 M solution in ethyl ether) to give 75 mg of awhite solid (48%). ESI-MS calculated for C₂₅H₃₄F₃N₃O₃: 481.26; found 482[M+H]⁺.

EXAMPLE 92

The product from Example 91 (11 mg, 0.021 mmol) was combined withformalin (37% aqueous solution, 17 μL, 0.21 mmol),N,N-diisopropylethylamine (8 μL, 0.05 mmol) and 4 Å powdered molecularsieves (20 mg) in 5 mL of dichloromethane. The mixture was stirred atroom temperature for 30 min before sodium triacetoxyborohydride (22 mg,0.10 mmol) was added. The reaction was stirred at room temperature for 1h before being filtered through celite, washed with saturated sodiumbicarbonate and brine, and dried over Na₂SO₄, filtered and concentratedunder reduced pressure. The product was purified by preparative TLC(silica gel, 0.5% NH₄OH/4.5% methanol/95% dichloromethane) and convertedto its hydrochloride salt by the addition of hydrogen chloride (2 Msolution in ethyl ether) to give 6.4 mg of a white solid (58%). ESI-MScalculated for C₂₆H₃₆F₃N₃O₂: 495.27; found 496 [M+H]⁺.

EXAMPLE 93

Intermediate 23 (37 mg, 0.079 mmol) was combined with Intermediate 26(10 mg, 0.079 mmol), N,N-diisopropylethylamine (43 μL, 0.25 mmol), 4 Åpowdered molecular sieves (50 mg), and sodium triacetoxyborohydride (50mg, 0.24 mmol) in 5 mL dichloromethane. The reaction mixture was stirredat room temperature for 24 h before being filtered through celite,washed with saturated sodium bicarbonate and brine, and dried overNa₂SO₄, filtered and concentrated under reduced pressure. The productwas purified by preparative TLC (silica gel, 0.5% NH₄OH/4.5%methanol/95% dichloromethane) and converted to its hydrochloride salt bythe addition of hydrogen chloride (2 M solution in ethyl ether) to give18 mg of a white solid (45%). ESI-MS calculated for C₂₄H₂₉F₆N₃O₂:505.22; found 506 [M+H]⁺.

EXAMPLE 94

Intermediate 24 (70 mg, 0.16 mmol) was combined with Intermediate 26 (20mg, 0.16 mmol), N,N-diisopropylethylamine (72 μL, 0.42 mmol), 4 Åpowdered molecular sieves (100 mg), and sodium triacetoxyborohydride(165 mg, 0.78 mmol) in 5 mL dichloromethane. The reaction mixture wasstirred at room temperature for 1.5 h and then was placed in therefrigerator over the weekend before being filtered through celite,washed with saturated sodium bicarbonate and brine, and dried overNa₂SO₄, filtered and concentrated under reduced pressure. The productwas purified by preparative TLC (silica gel, 0.5% NH₄OH/4.5%methanol/95% dichloromethane) and converted to its hydrochloride salt bythe addition of hydrogen chloride (2 M solution in ethyl ether) to give24 mg of a white solid (29%). ESI-MS calculated for C₂₄H₂₉F₆N₃O₃:521.21; found 522 [M+H]⁺.

EXAMPLE 95

Intermediate 16 (68 mg, 0.016 mmol) was combined with Intermediate 26(19 mg, 0.16 mmol), N,N-diisopropylethylamine (82 μL, 0.48 mmol), 4 Åpowdered molecular sieves (100 mg), and sodium triacetoxyborohydride(170 mg, 0.80 mmol) in 5 mL dichloromethane. The reaction mixture wasstirred at room temperature for 3 days before being filtered throughcelite, washed with saturated sodium bicarbonate and brine, and driedover Na₂SO₄, filtered and concentrated under reduced pressure. Theproduct was purified by preparative TLC (silica gel, 0.5% NH₄OH/4.5%methanol/95% dichloromethane) to give 2 diastereomers. The top spot andthe bottom spot where both converted to their hydrochloride salts by theaddition of hydrogen chloride (2 M solution in ethyl ether) to give 17mg and 5 mg respectively. Top Spot: ESI-MS calculated for C₂₄H₃₂F₃N₃O₃:467.24; found 468 [M+H]⁺. Bottom Spot: ESI-MS calculated forC₂₄H₃₂F₃N₃O₃: 467.24; found 468 [M+H]⁺.

EXAMPLE 96

To a solution of Intermediate 27 (20 mg, 0.057 mmol) in dichloromethane(10 mL) was added (3R,4R)₄-aminotetrahydrothiophene-3-ol 1,1-dioxide (17mg, 0.11 mmol), 4 Å powdered molecular sieves (50 mg) and sodiumtriacetoxyborohydride (60 mg, 0.28 mmol). The resulting reaction mixturewas stirred at room temperature for 3 days before being diluteddichloromethane and washed with saturated aqueous sodium bicarbonate andbrine. The organic layer was dried over sodium sulfate, filtered andconcentrated under reduced pressure. The resulting crude material waspurified by preparative TLC (4.5% methanol/0.5% NH₄OH/95%dichloromethane) to give the 2 desired single stereoisomers. Higherband: HPLC-MS calculated for C₂₂H₃₀F₃N₃O₄S: 489.19; found 490 [M+H]⁺.Lower band: HPLC-MS calculated for C₂₂H₃₀F₃N₃O₄S: 489.19; found 490[M+H]⁺.

EXAMPLE 97

This compound was prepared as detailed in Example 1 using2-methoxy-cyclohexanone instead of tetrahydro-4H-pyran-4-one. The fourisomers on the cyclohexane ring were resolved on a preparative chiral ODcolumn (5/95, ethanol/Hexanes). LC-MS for C₂₅H₃₆F₃N₃O₂ calculated:467.28, found: 468.25 [M+H]⁺.

EXAMPLE 98

This compound was prepared as detailed in Example 1 using2-methyl-cyclohexanone instead of tetrahydro-4H-pyran-4-one. The cis andtrans racemate in respect to the cyclohexane ring were resolved on apreparative chiral AD column (2/98, ethanol/Hexanes). LC-MS forC₂₅H₃₆F₃N₃O calculated: 451.28, found: 452.35 [M+H]⁺.

EXAMPLE 99

This compound was prepared as detailed in Example 30 using cyclohexanoneinstead of tetrahydro-4H-pyran-4-one. LC-MS for C₂₄H₃₄F₃N₃ calculated:453.26, found: 454.3 [M+H]⁺.

EXAMPLE 100

This compound was prepared as detailed in Example 30 using2-methoxy-cyclohexanone instead of tetrahydro-4H-pyran-4-one. LC-MS forC₂₅H₃₆F₃N₃O₃ calculated: 4834.27, found 484.3 [M+H]⁺.

EXAMPLE 101

Following the procedure described for Example 12 but using Intermediate31 instead of tetrahydro-4H-pyran-4-one afforded Example 101 as amixture of 4 diastereomers. Chiral separation on the AD column elutingwith isopropanol/heptane (8%) afforded the 4 resolved diastereomers.LC-MS for C₂₅H₃₆F₃N₃O₂ calculated 467.28 found 468.25 [M+H]⁺.

EXAMPLE 102

Following the procedure described for Example 12 but using ethyl4-oxocyclohexane-1-carboxylate instead of tetrahydro-4H-pyran-4-oneafforded Example 105 as mixture of 2 diastereomers. Chiral separation onthe OD column eluting with ethanol/heptane (15%) afforded the 2 resolveddiastereomers. LC-MS for C₂₇H₃₈F₃N₃O₃ calculated 509.29 found 510.4[M+H]⁺.

EXAMPLE 103

A solution of the diastereomeric esters in Example 102 (45 mg, 0.088mmol) in methanol (1.0 mL) was treated with an aqueous solution of LiOH.H₂O (10 mg, 2.4 mmol) and the mixture was stirred at 50° C. overnight.The volatiles were evaporated and the product purified by reverse phaseHPLC to afford Example 82. LC-MS for C₂₅H₃₄F₃N₃O₃ calculated 481.26found 482.35 [M+H]⁺.

EXAMPLE 104

Following the procedure described for Example 12 but using ethyl3-oxocyclohexane-1-carboxylate instead of tetrahydro-4H-pyran-4-oneafforded Example 104 as mixture of 2 diastereomers. LC-MS forC₂₇H₃₈F₃N₃O₃ calculated 509.29 found 510.4 [M+H]⁺.

EXAMPLE 105

A solution of the diastereomeric esters from Example 104 (65 mg, 0.13mmol) in methanol (1.5 mL) was treated with an aqueous solution of LiOH(20 mg, 0.48 mmol) and the mixture was stirred at 50° C. overnight. Thevolatiles were evaporated and the product was purified by reverse phaseHPLC. LC-MS for C₂₅H₃₄F₃N₃O₃ calculated 481.56 found 482.5 [M+H]⁺.

EXAMPLE 106

Following the procedure described for Example 12 but using ethyl3-(4-oxocyclohexyl) propanoate instead of tetrahydro-4H-pyran-4-oneafforded Example 106 as mixture of 2 diastereomers. LC-MS forC₂₉H₄₂F₃N₃O₃ calculated 537.32 found 538.5 [M+H]⁺.

EXAMPLE 107

A solution of the diastereomeric esters from Example 106 (75 mg, 0.14mmol) in methanol (1.5 mL) was treated with an aqueous solution of LiOH(25 mg, 0.60 mmol) and the mixture stirred at 50° C. overnight. Thevolatiles were evaporated and the product was purified by reverse phaseHPLC. LC-MS for C₂₇H₃₈F₃N₃O₃ calculated 509.29 found 510.5 [M+H]⁺.

EXAMPLE 108

The procedure described in Step A, Intermediate 19 was followed usingIntermediate 34 instead of Intermediate 8. LC-MS for C₂₈H₄₃N₃O₃calculated 469.33 found 470.3 [M+H]⁺.

A solution of the product from Step A in ethyl acetate at 0° C. wastreated with a saturated solution of HCl in ethyl acetate and themixture was stirred for 2 h. The volatiles were evaporated in vacuo toafford the title product as a white foam. LC-MS for C₂₃H₃₅N₃O calculated369.28 found 370.5 [M+H]⁺.

Following the procedure described for Example 12 but using the productfrom Step B instead of Intermediate 29 afforded Example 108. LC-MS forC₂₈H₄₃N₃O₂calculated 453.34 found 454.4 [M+H]⁺.

EXAMPLE 109

Starting from 0.235 g of Intermediate 35 and following the procedureoutlined for Intermediate 19, Step A gave 0.242 g of the title compound.LC-MS for C₂₇H₄₁N₃O₃ calculated 455.3 found 400.3 [M+H−56]⁺.

A solution of the product from Step A in ethyl acetate at 0° C. wastreated with a saturated solution of HCl in ethyl acetate and themixture was stirred for 2 h. The volatiles were evaporated in vacuo toafford 0.240 g of the title product. LC-MS for C₂₂H₃₃N₃O calculated355.2 found 356.3 [M+H]⁺.

Following the procedure described for Example 12 and starting from theintermediate prepared in Step B (0.1 g, 0.2 mmol) afforded 0.1 g ofExample 109 as its HCl salt. LC-MS for C₂₇H₄₁N₃O₂calculated 439.3 found440.4 [M+H]⁺.

EXAMPLE 110

Following the procedure described for Example 12 and starting fromIntermediates 19 (0.1 g, 0.2 mmol) and 32 afforded 0.0031 g of Example110 as mixture of diastereomeric compounds HCl salts. LC-MS forC₂₆H₃₆F₃N₃O₄ calculated 511.2 found 512.3 [M+H]⁺.

While the invention has been described and illustrated with reference tocertain particular embodiments thereof, those skilled in the art willappreciate that various adaptations, changes, modifications,substitutions, deletions, or additions of procedures and protocols maybe made without departing from the spirit and scope of the invention.For example, effective dosages other than the particular dosages as setforth herein above may be applicable as a consequence of variations inthe responsiveness of the mammal being treated for any of theindications with the compounds of the invention indicated above.Likewise, the specific pharmacological responses observed may varyaccording to and depending upon the particular active compounds selectedor whether there are present pharmaceutical carriers, as well as thetype of formulation and mode of administration employed, and suchexpected variations or differences in the results are contemplated inaccordance with the objects and practices of the present invention. Itis intended, therefore, that the invention be defined by the scope ofthe claims which follow and that such claims be interpreted as broadlyas is reasonable.

1. A compound selected from:

or a salt thereof.
 2. The compound:

or a salt thereof.
 3. The compound:

or a salt thereof.
 4. The compound:

or a salt thereof.
 5. The compound:

or salt thereof.